Detection Of Mycoplasma Agalactiae Research Articles

Detection of Mycoplasma agalactiae in Ticks (Rhipicephalus bursa) Collected by Sheep and Goats in Sicily (South-Italy), Endemic Area for Contagious Agalactia.

The aim of this preliminary study was to investigate the presence of Mycoplasma agalactiae (Ma) or other Contagious Agalactia (CA) causative organisms, in hard ticks infesting milking sheep and goats in endemic areas for CA in Sicily (South-Italy). Although there is accumulating evidence to support the role of ticks in the transmission of blood-borne haemoplasmas, information regarding their role in the transmission of CA, remains scarce. Ticks (n = 152) were collected from 25 lactating sheep and goats from three farms with previous outbreaks of CA. Microbiological and biomolecular, as well as serological analysis were performed on milk, tick, and serum samples, respectively. Rhipicephalus bursa species predominated, comprising 84.8% of the sampled ticks. Mycoplasma-like colonies were isolated from 5/56 (8.9%) tick pools and were identified as Ma by specific PCR and 16S rRNA gene sequencing. Unexpectedly, the organism was isolated from R. bursa ticks recovered only from animals whose milk tested negative for the pathogen. This preliminary demonstration suggests the potential role for ticks to act as a reservoir for the organisms, with potential involvement in the spread and maintenance of CA. Further work is required to determine the location of the organisms within the body of the ticks and to assess transmission potential.

Detection of Mycoplasma agalactiae by In-situ Hybridization and Characterization of Inflammatory Infiltrates by Immunohistochemistry in Sheep Udders

Detection of Mycoplasma agalactiae by In-situ Hybridization and Characterization of Inflammatory Infiltrates by Immunohistochemistry in Sheep Udders

Isolation and Detection of Mycoplasma agalactiae from Semen Samples of Goats.

Contagious agalactia (CA) is a highly infectious disease of goats and sheep, and is a form of Mycoplasmosis, which is usually enzootic. Since Mycoplasma agalactiae (M. agalactiae) is the main cause of this disease in goats, the aim of this study was to isolate and detect M. agalactiae from semen of goat bucks. Thirty-nine semen samples were collected from goat bulks, and all samples were cultured in PPLO broth medium supplemented for M. agalaciae isolation. The bacteria DNAs were extracted from clinical samples and the PCR assay was applied to detect Mycoplasma genus and M. agalactiae species using specific primers, which amplified a 163bp fragment in 16SrRNA gene and a 375bp fragment in lipoprotein gene. The PCR evaluations were performed for both the clinical samples and the cultures. Out of the 39 samples, 29 (74.3%) of the cultures were shown positive and typical Mycoplasma colonies grew on PPLO agar, which could be considered as the diagnostic method. In addition, 38 (97.4%) samples had positive PCR results for Mycoplasma genus and six (15.3%) of the samples were shown to be positive using PCR for M. agalactiae as the diagnostic method. In the present study, M. agalactiae was detected in semen of goat bulks for the first time in Iran. Therefore, it is recommended to concern semen as one of the significant sources for this pathogen and the possibility for transmission to the female goats through semen is highlighted. Moreover, presence of this microorganism in semen could be involved in infertility of goat population.

Detection of Mycoplasma agalactiae by culture and polymerase chain reaction (PCR) methods from Iranian goats

Mycoplasma agalactiae is the etiological agent of contagious agalactia that is a serious disease affecting sheep and goats. It is characterized by mastitis and subsequent failure of milk production, arthritis and keratoconjunctivitis. The aim of the study was to detection of M. agalactiae by culture and polymerase chain reaction (PCR) methods from infected Iranian goats. For detection, a total of 57 samples were taken from conjunctiva (n=11), joint exudates (n=35) and milk secretion (n=11). After enrichment in PPLO broth media, identification of the Mycoplasma species was carried out by cultured and PCR method. Of the 57 samples in 9 samples (16%) fried egg colony appeared on the agar media. The PCR with mycoplasmal 16S rRNA was applied for detection of a variety of Mycoplasmaspecies. PCR identification of genus was successful in 31 isolates (54%) and showed specific amplicon at 163 bp. From this positive samples, 19 isolates were examined (61%) were positive for M. agalactiae that showed specific amplicon at 375 bp. Of the total samples, only in 3 samples both culture and genus PCR tests were positive and in 20 samples were negative. Whereas in 6 PCR negative samples Mycoplasma colony appeared on the agar media and this colonies did not observed in 28 PCR positive samples. This paper reported detection of M. agalactiae from goats for the first time in Iran. PCR can be used as trusty and supersede test in the detection of M. agalactiae from affected goats. Among different collecting sites, milk secretion samples are suitable for PCR detection of M. agalactiae. Key words: Mycoplasma agalactiae, goat, polymerase chain reaction, culture, Iran.

A real-time polymerase chain reaction assay for the detection of Mycoplasma agalactiae

AIM: To develop a real-time PCR for the detection of Mycoplasma agalactiae, using PCR primers targeting the ma-mp81 gene. METHODS: A group of 15 M. agalactiae isolates, 21 other Mycoplasma spp. isolates and 21 other bacterial isolates was used in evaluation of the assay. RESULTS: All M. agalactiae isolates were detected by the assay and none of the non-target isolates was amplified. The analytical detection limit of the assay was 10 fg of purified genomic DNA and 104 cfu/ml milk inoculated with M. agalactiae. When applied to goat-milk samples collected from three herds free of M. agalactiae infection, the assay had a specificity of 100%. CONCLUSIONS: The assay would be useful in a diagnostic laboratory, providing specific, sensitive and rapid detection of M. agalactiae.

Detection of Mycoplasma agalactiae by Polymerase Chain Reaction in Jordanian Sheep and Goat Herds

The aim of the study was to ascertain whether sheep and goats from selected Jordanian herds were infected with Mycoplasma agalactiae, the most common aetiological agent of contagious agalactia of sheep and goats. All examined animals showed clinical signs of disease at the time of sample collection. The group included 35 animals, 15 sheep and 20 goats. For microbiological examination, a total of 107 swabs were taken from conjunctival, nasal, vaginal or preputial mucosae and from the external auditory canal. Identification of the species isolated was carried out by a polymerase chain reaction. Of the 35 animals, 21 (4 sheep and 17 goats) tested positive for Mycoplasma agalactiae. These results confirmed our assumption that this mycoplasma species is present in Jordanian herds and, for the first time, provided evidence that contagious agalactia of sheep and goats occurs in Jordan.

Detection of Mycoplasma agalactiae in sheep milk samples by polymerase chain reaction

We developed a simple and rapid method for DNA extraction from sheep milk to use for polymerase chain reaction (PCR) diagnosis of Mycoplasma agalactiae. We tested 357 samples from 21 newly infected flocks (group 1) and 87 samples from 8 flocks infected in the past (group 2). PCR results were compared with those of conventional culture. By PCR we detected 175 positives in group 1, while by culture we detected only 153. Milk samples from group 2 were negative, both by PCR assay and by culture. Our PCR is much faster than culture and reduces the time required for diagnosis from several days to 5 h. The method could be used for the routine diagnosis of contagious agalactia caused by Mycoplasma agalactiae.