Isolation And Molecular Detection of Mycoplasma Mycoides Cluster In Goats With Clinical Signs Of Contagious Agalactia In Kerman Province, Iran (original) (raw)
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Investigation of Mycoplasma agalactiae in sheep in Kurdistan province by PCR
archives of razi institute, 2015
Contagious agalactia (CA) is a serious disease syndrome of sheep and goats that is characterized by mastitis, arthritis, keratoconjunctivitis and, occasionally, abortion. Mycoplasma (M.) agalactiae is the main cause of the disease in sheep and goats. The aim of the present study was to detect M. agalactiae in conjunctival, synovial fluid, nasal, ear and milk samples in sheep herds with or without CA sign in Kurdistan province. One hundred and seventy three samples analyzed were taken from sheep herds reared in a CA endemic area. Of the samples tested, 130 were positive by PCR for Mycoplasma spp. and of these, 19 showed a positive result for M. agalactiae . M. agalactiae was detected in conjunctival (7/59), synovial fluid (1/8) and milk (11/21) samples. Ear and nasal swap samples were free of M. agalactiae . The results of 60 samples in sheep without signs of CA syndrome showed that Mycoplasma spp. and M. agalactiae were detected in 25 and 6 samples, respectively. Our findings indica...
2017
Contagious agalactia is an infectious syndrome of sheep that is characterized by mastitis with reduction of milk production, arthritis, abortion, and keratoconjunctivitis. The disease is rapidly spread by the contact of the infected animals with the healthy ones. Domestic sheep and goats of both sexes can be infected at an equivalent frequency. Most of the researchers use culture and molecular methods for the isolation and identification of Mycoplasma. Mycoplasma agalactiae is the main cause of the disease in sheep. The aim of this study was to isolate and identify M. agalactiae by using culture and polymerase chain reaction (PCR) assay in the sheep herds in Guilan province, Iran. A total of 71 specimens were collected from seven sheep herds with clinical signs of agalactia disease. All of the seven sheep herds (100%) were positive either in PPLO agar or Mycoplasma PCR test. Out of the 71 specimens, 50 (70.4%) cases were positive; however, 21 (29.6%) samples were negative. Furthermo...
Comparison of PCR tests for the detection of Mycoplasma agalactiae in sheep and goats
TURKISH JOURNAL OF VETERINARY AND ANIMAL SCIENCES, 2016
Introduction Mycoplasma species cause several diseases in small ruminants, which often manifest with clinical signs including mastitis, arthritis, keratitis, pneumonia, and septicaemia (1). Contagious agalactia (CA) is caused by Mycoplasma agalactiae, while Mycoplasma mycoides subsp. capri, Mycoplasma capricolum subsp. capricolum, and Mycoplasma putrefaciens cause diseases similar to CA, all of which have been highlighted by the World Organisation for Animal Health (1) due to the significant economic impact associated with reduced milk production and quality (2). CA is widespread and occurs in many countries bordering the Mediterranean in Europe, Asia, and Africa (2,3). Additionally, CA has been endemic in several regions of Turkey for many years. The presence and distribution of mycoplasma diseases in sheep and goats in Turkey has been explored previously (4-6). Mycoplasma species are fastidious organisms, which makes traditional diagnosis and identification by culture, biochemical, and serological methods difficult and time consuming. Therefore, molecular methods such as the polymerase chain reaction (PCR) are quicker and generally more sensitive than culture. However, with more than 120 different known mycoplasma species, a single molecular technique that can detect and identify the different species in one test differentiate has negated the need to develop 120 different PCR tests. PCR of the 16S rRNA gene followed by denaturing gradient gel electrophoresis (PCR/DGGE) determines the genetic diversity of microorganisms, as it can theoretically detect single base mutation in DNA and also identify the majority of small ruminant mycoplasmas (7,8). This method uses G-C rich clamped primers based on the 16S rDNA gene and results in amplicons that are separated in a urea and formamide gradient polyacrylamide gel heated at 60 °C (9). The PCR/DGGE method can detect and differentiate 67 mycoplasma species of veterinary importance including causative bacteria of CA (1,7,10). PCR/DGGE can be applied directly to clinical samples or cultured isolates. It can also detect new mollicute species and detect and identify mixed cultures (11). In this study, we compared two M. agalactiae-specific PCR methods (12,13) with PCR of the 16S rRNA gene followed by denaturing gradient gel electrophoresis (PCR/DGGE) (7) using samples collected from sheep and goats. These experiments also indicated the presence of Mycoplasma species and their association with clinical and subclinical signs of CA in sheep and goats.
Detection of Mycoplasma agalactiae in Small Ruminants of Southeast Iran
2017
Agalactia is an infectious and contagious disease of small ruminants caused by Mycoplasma agalactiae (M. agalactiae). Although different microorganism strains contribute to this disease, M. agalactiae is known as the most prominent causative agent. Therefore, this study aimed to investigate the rate of M. agalactiae involvement in contagious agalactia in the southeast region of Iran. Sampling was performed from milk, conjunctiva, ear lesions, and joints exudate of suspicious sheep and goat flocks according to the reports of Iran Veterinary Organization. The presence of Mycoplasma and its species, namely M. agalactiae, was evaluated through microbial culture and polymerase chain reaction (PCR) techniques. The detected microorganisms were confirmed to be Mycoplasma and M. agalactiae by the PCR amplification of 16S rRNA and lipoprotein target genes. According to the findings of present study, 14.8% and 36.0% of the samples were diagnosed as positive for Mycoplasma by culture and PCR, r...
Detection of Mycoplasma agalactiae by Polymerase Chain Reaction in Jordanian Sheep and Goat Herds
Acta Veterinaria Brno, 2007
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. Mycoplasma agalactiae, sheep, goats, Jordan, polymearase chain reaction Contagious agalactia of sheep and goats is a serious infectious disease affecting small ruminants. It is characterised by arthritis, keratoconjunctivitis, pneumonia and, in females, mastitis and occasional abortion. It may be caused by any of the five mycoplasma species associated with this disease, namely, Mycoplasma agalactiae, Mycoplasma mycoides subsp. mycoides LC (large colony), Mycoplasma capricolum subsp. capricolum, Mycoplasma mycoides subsp. capri and Mycoplasma putrefaciens (Nicholas 1996; Sarris 1996; Bergonier et al. 1997; Bölske 1994). However, Mycoplasma agalactiae (M. agalactiae) is still regarded, particularly in sheep, as the "classical" aetiological agent of contagious agalactia (Bergonier et al. 1997). Mycoplasma putrefaciens is found only in goats. However, other animal species, such as cattle, camels or small wild ruminants, can also be infected (Perrin et al. 1994). Contagious agalactia is at present an important disease in countries with intensive small animal husbandry, such as the Mediterranean region and the Balkan Peninsula, Western
African Journal of Microbiology Research, 2013
Mycoplasma agalactiae is the etiological agent of contagious agalactia, which is the first isolated mycoplasma from sheep and goats. The bacteria have been detected using polymerase chain reaction (PCR) method in Iranian goats, but there has been no investigation of the molecular characterization of contagious agalactia agents in goats in Iran. Several studies have also characterized M. agalactiae strains. The aim of this study was to analyze the DNA sequence of M. agalactiae isolates from Iranian goats, and compare it with the other Iranian isolates and vaccinal strains obtained from GenBank. Therefore, PCR with a mycoplasmal cytoplasmic membrane protein 80 KDa (P80) gene was applied to Iranian isolates and the PCR product was purified and sequenced. After the computer alignment of these gene sequences was done, the next process was comparing the result with the specific species of M. agalactiae deposited in GenBank. However, a sequence identity matrix is represented (Bio Edit, Clustal W) and a phylogenetic tree is built by the neighbor-joining method. 1000 bootstrap simulations were given at the tree's branching points and represent these points' phylogenetic relationships. Sequence analysis of different M. agalactiae showed significant variability among the field isolates. In comparison with vaccine strains, which are used in Iranian goat herds, characterized isolates under this study has a poor homology.
Investigation of Mycoplasma agalactiae by molecular methods in goat bucks in West of Iran
The presence of asymptomatic carriers of Mycoplasma agalactiae in goat buck populations, and the internal presence of this microorganism in some of these animals, indicated that the ability of this micro-organism captive in contagious agalactiae to colonize in other locations such as the respiratory system or genitalia. This study was designed to detect the excretion Mycoplasma agalactiae in goat bucks reared in Iran, where the disease is considered endemic. A total of 196 samples which included semen (n=49), conjunctiva (n=49), nasal (n=49) and ear (n=49) of goat bucks were collected from July to September 2013. To avoid the introduction of goat bucks carrying Mycoplasma organisms sub clinically in samples were obtained from all candidate goat bucks for PCR, to detect the presence of Mycoplasma agalactiae. Of the samples tested, 31.12% were scored positive for Mycoplasma spp. and Mycoplasma agalactiae was detected in seventeen samples, conjunctival (6 positive sample), nasal (1 positive sample), ear (5 positive sample) and semen (5 positive sample). The results obtained in the present study confirm of Mycoplasma agalactiae in the semen, conjunctival, nasal and ear in goat bucks without clinical symptoms of contagious agalactiae infection. The presence of this agent in male goat is considered very risky in terms of reproductive disorders and contagious agalactiae outbreaks in endemic areas in Iran. This is the first report of Mycoplasma agalactiae in goat bucks without contagious agalactiae signs in Iran.
2018
Respiratory tract infections are the major cause of economic loss in goats. Mycoplasma agalactiae acts as an important etiological agent in mastitis, arthritis, kerato-conjuctivitis, pleuropneumonia and septicaemia syndrome (MAKePS). This paper reports a case study regarding the outbreak of respiratory disease associated with M. agalactiae in a herd of goats in Thrissur district, Kerala. The goats were presented with symptoms of respiratory distress, conjunctivitis, lethargy and reduced production. Nasal swabs were collected and dispensed in modified Hayfick’s broth without crystal violet. After incubation at 37°C for about 72 h., DNA was isolated from the broth and subjected to Mycoplasma genus specific PCR, which revealed amplicons of 280 bp confirming the genus. Also, the solid medium which was inoculated with filtered drops of incubated broth presented typical fried egg like colonies after five days of incubation and the isolates were identified as Mycoplasma by PCR. The species...
Al-Qadisiyah Journal of Veterinary Medicine Sciences, 2016
Mastitis is the main signs of contagious agalactia syndrome in sheep and goats, which is caused by Mycoplasma agalactiae and induces heavy economic losses. The aim of this study was to isolate and identify M. agalactia from mastitic lactating ewes at Al-Muthana province. Clinical mastitic milk collected from infected animals. The collected samples were transferred in a transport medium which contained PPLO broth, horse serum and yeast extract at cool conditions to the lab. All the samples were carried out by PCR and cultured on PPLO agar too. The results showed that, from the total of 150 mastitic milk samples, 40% was positive by culture for the growth of mycoplasma. Out of 60 positive culture, there was 35(58.3%) positive for Mycoplasma spp. PCR and 4\35 (11.4%) were identified as Mycoplasma agalactiae by PCR. The lactating ewes with mycoplasmas mastitis exhibited non-significant variation in temperature, respiration and pulsation rates when compared with healthy ewes group.
Species of Mycoplasma causing contagious agalactia in small ruminants in Northwest Iran
Contagious agalactia (CA) is a serious disease of small ruminants that occurs in many countries, and is usually characterized by mastitis, arthritis, keratoconjunctivitis, pleuropneumonia, and septicemia. Mycoplasma agalactiae (Ma) is the main causative agent in sheep and goats but other pathogens including Mycoplasma mycoides subsp. capri (Mmc, which incorporates the former M. mycoides subsp. mycoides Large Colony type), Mycoplasma capricolum subsp. capricolum (Mcc), and Mycoplasma putrefaciens (Mp) might be involved. They are all usually associated with infections in goats and may cause similar clinical signs. A total of 116 sheep and 16 goats suffering from the acute form of the disease were included in this study. They were recruited following a number of outbreaks suspected to be CA in the Ardebil province of Iran. Milk, lachrymal or synovial fluid were collected exclusively from the affected animals in order to identify the pathogen involved. Of the 132 collected samples, 33 (25%) were positive for Mycoplasma species by culture in PPLO broth and agar. The polymerase chain reaction followed by denaturing gradient gel electrophoresis (PCR/DGGE) method identified 18 (12 sheep and 6 goats) of the 33 Mycoplasma positive samples with mixed Mycoplasma population. In particular, 25 Ma (47.2%), 23 Mp (43.4%), 4 Mcc (7.5%), and 1 Mmc (1.9%) were identified. This confirms that the several Mycoplasma species rather than the Ma only are in circulation, and are able to cause CA in sheep and goats in Iran. This is the first report on the isolation and identification of Mp, Mmc and Mcc in infected small ruminant flocks in Iran.