Candidatus Mycoplasma Haemolamae Research Articles

Molecular survey of hemoplasmas in asymptomatic vicunas (Vicugna vicugna) from the Pampa Galeras Bárbara D'Achille National Reserve in Peru

Hemotrophic mycoplasmas (hemoplasmas) are epierythrocytic bacteria that infect wild and domestic animals, and can cause anemia in some of them. They are considered emerging and zoonotic pathogens, causing serious health problems in wildlife. Candidatus Mycoplasma haemolamae is the only species of hemoplasma that infects domestic South American camelids (alpacas and llamas), with limited studies in wild camelids. Therefore, the objective of this study was to determine the prevalence of Candidatus M. haemolamae in vicunas (Vicugna vicugna) from the Pampa Galeras National Reserve, located in the Ayacucho region of Peru, using molecular diagnosis. For this, blood samples from 79 vicunas were collected, which were molecularly analyzed by partially amplifying the 16S ribosomal RNA gene of Mycoplasma sp. Fourteen vicunas (17.7 %) were positive for the molecular diagnosis of Mycoplasma sp. All PCR-positive products were sequenced and showed more than 99 % identity with Candidatus M. haemolamae. Statistical analysis showed that tick-infested vicunas had 6.10 odds of presenting Candidatus M. haemolamae compared with tick-free vicunas. Sex and age were not associated with Candidatus M. haemolamae infections. This is the first report of hemoplasmas in vicunas, a wild South American camelid, demonstrating that the pathogen can have both a domestic and a wild life cycle. Future studies are necessary to know the current situation of this pathogen in domestic and wild camelids from other locations in Peru.

Molecular survey and genetic characterization of ‘Candidatus Mycoplasma haemolamae’ in llamas (Lama glama) and alpacas (Vicugna pacos) from Southern Chile.

This study aimed to perform a molecular survey and identification of hemotropic Mycoplasma spp. in domestic South American Camelids from Southern Chile. Conventional PCR (cPCR) for hemotropic Mycoplasma spp. based on 16S rRNA gene (620bp fragment) was performed in 87 EDTA-blood samples taken from 48 llamas (Lama glama) and 39 and alpacas (Vicugna pacos) from to Temuco, La Araucanía region and Valdivia, Los Rios region, Southern Chile. 16S rRNA hemotropic Mycoplasma PCR-positive were sequenced for species identification, phylogenetic and haplotype analyses, and further tested by cPCR targeting a fragment (160-210 bp) of the RNaseP (rnpB) gene. Based upon 16S rRNA cPCR results, the overall hemotropic Mycoplasma spp. occurrence in Southern camelids was 9.2% (8/87 [95% CI (4.0–17.3%)]), with five positive alpacas (12.8%; 5/39 [95% CI (4.3–27.4%)]) and three llamas (6.3%; 3/48 [95% CI (1.7–17.2%)]). All 16S rRNA PCR-positive samples were negative for the rnpB gene. Obtained 16S sequences presented high identity (99-100%) by BLASTn analysis to ‘Candidatus Mycoplasma haemolamae’ from an alpaca in the United Kingdom. Phylogenetic and haplotype analyses of the 16s rRNA gene showed high similarity among ‘Candidatus M. haemolamae’ sequences of this study and the ones from North America, Europe, and Asia evidencing a low diversity of Chilean samples, with only one haplotype detected (#1). Haplotype #1 from South American Camelids in Chile was worldwide distributed and observed in North America, Europe, and Asia. ‘Candidatus M. haemolamae’ detected for the first time in South American camelids in Southern Chile had low diversity and was worldwide spread.

Trypanosoma evansi and “Candidatus Mycoplasma haemolamae” Co-Infection in One-Humped Camel (Camelus dromedarius) from the Northwest of Iran: A Case Report

A 4-year-old male one-humped camel (Camelus dromedarius) was referred to Veterinary Teaching Hospital of Urmia University, Iran in 2017 with anorexia, weakness, depression and pale mucosa. Decreased red blood cell count, packed cell volume and hemoglobin concentration were detected by complete blood cell count. In Giemsa-stained peripheral blood smears Trypanosoma spp. trypomastigotes scattered between erythrocytic spaces and Mycoplasma-like organisms were observed attached to the surface of erythrocytes. Species-specific PCR assay confirmed T. evansi and Candidatus Mycoplasma haemolamae (CMhl) co-infection. Administration of diminazene aceturate, oxytetracycline 20%, flunixin meglumine and phosphorus-vitamin B12 were not effective in treatment. Hemoplasmosis should be considered as an important differential diagnosis of conditions associated with hemolytic anemia in camel.

Genetic characterization and phylogenetic analysis of hemotrophic mycoplasmas in camel (Camelus dromedarius)

So far, a number of hemotrophic mycoplasma (HM) species have been reported from different animals, mostly based on the 16S rDNA gene sequence analysis. Although previous studies have described Candidatus Mycoplasma haemolamae from alpaca (Lama pacos), llama (Lama glama), and vicuna (Lama vicugna) in Camelidae, data on the causative agent(s) of hemotrophic mycoplasmosis or hemoplasmosis are insufficient in dromedary camel (Camelus dromedarius). The purpose of the present molecular study was to investigate hemotrophic mycoplasmosis in Iranian dromedary camel. Accordingly, a total number of 100 EDTA-anticoagulated blood specimens were randomly collected via the jugular vein from camels. In the present study, partial sequence data of the 16S rDNA was employed as a molecular approach for genetic characterization and phylogenetic analysis of the Iranian strains of hemotrophic mycoplasma in camel. The phylogenetic analysis revealed that one species belonged to the “haemominutum group” within the cluster of hemoplasmas (the former genus Eperythrozoon), the other phylogenetic analysis found species belonging to the “haemofelis group” with the hemoplasma cluster (representing the former genus Haemobartonella). Out of the 100 tested blood samples, 2 (2.0%) were found positive by the PCR assay. The phylogenetic tree inferred from 16S rDNA sequences showed that examined camels were naturally infected with two distinct species of hemotrophic mycoplasmas. The first strain was phylogenetically placed into an undescribed hemotrophic mycoplasma group with a new species of Mycoplasma sp. isolated from a pig in China with 99% genetic similarity. However, the second strain seemed basal to a large lineage of Mycoplasma suis with a low sequence similarity of 93.5–95.9%. In summary, this is the first molecular-based study to detect the presence of two genetically different hemotrophic mycoplasma strains in dromedary camel that are phylogenetically linked to two pathogenic swine species.

Prevalence of Candidatus Mycoplasma haemolamae, bovine viral diarrhoea virus, and gastrointestinal parasitism in a sample of adult New Zealand alpaca (Vicugna pacos)

AIM: To determine the prevalence of infection with Candidatus Mycoplasma haemolamae (Mhl), antibodies to bovine viral diarrhoea virus (BVDV), and BVDV antigen, and the prevalence of animals with elevated faecal nematode egg counts (FEC) in a sample of adult New Zealand alpaca (Vicugna pacos).METHODS: Blood samples were obtained from 175 alpaca, collected from 15 farms around New Zealand, and from 31 samples sent to a diagnostic laboratory for routine haematology. Blood smears (n=170) were examined microscopically for the presence of haemoplasma, and DNA was extracted from whole blood (n=206) for real-time PCR testing for Mhl. Packed cell volume (PCV) was determined for 193 samples. Serum samples (n=195) were tested for BVDV antibody using ELISA, and for BVDV antigen using a real-time PCR assay. Faecal samples were collected from 143 animals; FEC were measured, and samples pooled for larval culture.RESULTS: No haemoplasma organisms were present on blood smear examination. Of the 206 blood samples, two (from the same farm) were positive for Mhl by real-time PCR testing, giving a prevalence of infection with Mhl of 0.97%. Of the 195 serum samples tested, four (2.1%) were positive for antibodies to BVDV; animals with BVDV antibodies were from 3/15 (20%) farms, none of which farmed cattle. None of the serum samples were positive by PCR for BVDV antigen. The median FEC was 50 epg (min 0, max 4,700), with 55/143 (38.5%) samples having 0 epg, and 33/143 (23.1%) having ≥250 epg. Haemonchus spp. were the most common nematodes present in faecal larval cultures from the North Island. Log10 FEC was negatively associated with PCV (p=0.02), and was higher in males than females (p<0.001), and in animals that were positive compared with negative for Mhl (p=0.022).CONCLUSIONS AND CLINICAL RELEVANCE: The number of alpaca infected with Mhl was low, as was the seroprevalence of BVDV. Gastrointestinal parasitism was, however, a common finding in this sample of New Zealand alpaca.

First detection of ‘Candidatus Mycoplasma haemolamae’ infection in alpacas in England

This is the first report of detection of Candidatus Mycoplasma haemolamae in alpacas in England. The primary case occurred in a three year-old male alpaca in the south-east of England...

Genome Sequence of “Candidatus Mycoplasma haemolamae” Strain Purdue, a Red Blood Cell Pathogen of Alpacas (Vicugna pacos) and Llamas (Lama glama)

We report the complete genome sequence of "Candidatus Mycoplasma haemolamae," an endemic red-cell pathogen of camelids. The single, circular chromosome has 756,845 bp, a 39.3% G+C content, and 925 coding sequences (CDSs). A great proportion (49.1%) of these CDSs are organized into paralogous gene families, which can now be further explored with regard to antigenic variation.

Progress towards less invasive veterinary surgery

ONE of the most remarkable areas of advance in medical science over the past two or three decades has been the development of methodologies for surgical and diagnostic procedures that...

First detection of ‘Candidatus Mycoplasma haemolamae’ infection in alpacas in England

This is the first report of detection of Candidatus Mycoplasma haemolamae in alpacas in England. The primary case occurred in a three year-old male alpaca in the south-east of England...

Novel hemotrophic mycoplasma identified in naturally infected California sea lions (Zalophus californianus)

The hemoplasmas are the trivial name for a group of erythrocyte-parasitizing, non-cultivable in vitro bacteria of the genus Mycoplasma that have been described in several mammalian hosts worldwide. This study is the first report of hemoplasmas in marine mammals. EDTA anticoagulated whole blood samples from 137 California sea lions (Zalophus californianus) and 20 northern elephant seals (Mirounga angustirostris) admitted to the Marine Mammal Center (Sausalito, CA; www.marinemammalcenter.org) or live captured in Oregon were collected during 2008. Hemoplasma-specific genomic DNA was detected in blood samples from 12.4% California sea lions tested using PCR. Hemoplasma PCR positive blood specimens also were tested in reverse transcription polymerase chain reaction (RT-PCR) using the hemoplasma-specific primers for the 16S and 23S rRNA genes. The RT-PCR showed the presence the hemoplasmal rRNA, strongly suggesting the presence of potentially viable hemoplasmas in the bloodstream of the animals. BLAST search and phylogenetic analysis of the 16S rRNA sequences of the hemoplasma from California sea lions revealed that the organism is a novel hemoplasma species with only 92.1% of its nucleotide similarity to the 16S rRNA gene of the previously described hemoplasma species of alpacas, Candidatus Mycoplasma haemolamae. Thus, due to low level of genetic similarity of the hemoplasma to other described hemoplasmas and the mammalian host in which the hemoplasma was detected we propose that this novel hemoplasma species has been given the provisional name Candidatus Mycoplasma haemozalophi sp. nov.

First detection of "Candidatus Mycoplasma haemolamae" in South American Camelids of Switzerland and evaluation of prevalence

Haemotrophic mycoplasmas (also known as haemoplasmas), small bacterias which parasite the surface of erythrocytes, have been described in several species. Recently, molecular methods were developed for the diagnosis of haemoplasma infection. The presented study describes the first detection and the investigation of prevalence of "Candidatus Mycoplasma haemolamae" in South American Camelids in Switzerland. A random sample of the latter population was tested for haemoplasma infections using real-time PCR. The infection was detected in 18.6% of the animals and was found both in indigenous and in imported camelids. Of the tested herds 39,1% harboured at least one animal positive for haemoplasmas in PCR. There was no difference in prevalence between male and female animals and llamas and alpacas, respectively. Furthermore, the prevalence of infection was not significantly different in diseased animals compared to healthy camelids. From the latter observation and the fact that the high prevalence was accompanied by an undetectable incidence, we concluded that the pathogenicity of "Candidatus Mycoplasma haemolamae" may be low.

Development and application of a real-time TaqMan ® qPCR assay for detection and quantification of ‘ Candidatus Mycoplasma haemolamae’ in South American camelids

Two alpacas from a herd in southwest Switzerland died for unknown reasons. Necropsy revealed chronic weight loss and pale mucous membranes. Infection with hemotropic mycoplasmas was suspected and subsequently confirmed by molecular methods. In order to investigate the epidemiological situation in this herd, a real-time TaqMan ® qPCR assay for the specific detection and quantification of hemoplasma infection in South American camelids was developed. This assay was based on the 16S rRNA gene and amplified ‘ Candidatus Mycoplasma haemolamae’ DNA, but not DNA from other hemoplasmas or non-hemotropic mycoplasma species. The lower detection limit was one copy/PCR, and the amplification efficiency was 97.4%. In 11 out of 24 clinically healthy herd mates of the two infected alpacas, ‘ Candidatus M. haemolamae’ infection was confirmed. No correlation was found between bacterial load and clinical signs or anemia. The assay described herein enables to detect and quantify ‘ Candidatus M. haemolamae’ and may be used in future studies to investigate the prevalence, pathogenesis and treatment follow-up of hemoplasma infections in South American camelids.

Use of a polymerase chain reaction assay to study response to oxytetracycline treatment in experimental Candidatus Mycoplasma haemolamae infection in alpacas

To develop a PCR assay for Candidatus Mycoplasma haemolamae (CMhl) infection in alpacas and use it to study the efficacy of oxytetracycline treatment and development of a subclinical carrier state. 8 healthy adult alpacas. Alpacas initially had negative results for CMhl in blood samples via PCR assay and were experimentally infected with CMhl; 4 were treated with oxytetracycline, and 4 were not treated. All were monitored regularly via PCR assay, blood smear examination, PCV, rectal temperature, and physical examination. At 6 months after treatment, all alpacas were immunosuppressed by administration of dexamethasone and tested for CMhl. 7 of 8 alpacas had positive PCR assay results 4 to 6 days after experimental infection. When organisms were detectable on a blood smear, they were seen 2 to 6 days after positive results of PCR assay. Infection was often associated with mild anemia that was usually transient. No alpacas became hypoglycemic. Oxytetracycline treatment was not associated with faster clearance of organisms or resolution of anemia, and 4 of 4 treated alpacas still had positive results of PCR assay when immunosuppressed 6 months later; 0 of 3 nontreated alpacas had positive results of PCR assay following immunosuppression. Transient fever was detected in 3 alpacas during immunosuppression. The PCR assay was more sensitive than blood smear examination for detection of infection. Clinical signs, anemia, and fever were not necessarily associated with infection. Oxytetracyline administration did not consistently clear CMhl infection. Although treated with oxytetracycline, infected alpacas remained chronic carriers.

'Candidatus mycoplasma haemodidelphidis' sp. nov., 'Candidatus mycoplasma haemolamae' sp. nov. and Mycoplasma haemocanis comb. nov., haemotrophic parasites from a naturally infected opossum (Didelphis virginiana), alpaca (Lama pacos) and dog (Canis familiaris): phylogenetic and secondary structural relatedness of their 16S rRNA genes to other mycoplasmas.

The 16S rRNA sequence of newly characterized haemotrophic bacteria in an opossum (Didelphis virginiana) and alpaca (Lama pacos) was determined. In addition, the 16S rRNA sequence of a haemotrophic parasite in the dog (Canis familiaris) was determined. Sequence alignment and evolutionary analysis as well as secondary structural similarity and signature nucleotide sequence motifs of their 16S rRNA genes, positioned these organisms in the genus Mycoplasma. The highest scoring sequence similarities were 16S rRNA genes from haemotrophic mycoplasma species (Haemobartonella and Eperythrozoon spp.). However, the lack of several higher-order structural idiosyncrasies used to define the pneumoniae group, suggests that these organisms and related haemotrophic mycoplasmas represent a new group of mycoplasmas. It is recommended that the organisms be named 'Candidatus Mycoplasma haemodidelphidis', 'Candidatus Mycoplasma haemolamae' and Mycoplasma haemocanis comb. nov., to provide some indication of the target cell and host species of these parasites, and to reflect their phylogenetic affiliation.