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Hemoplasmas and ticks in cattle from Somalia
Abstract
Hemotropic mycoplasmas (hemoplasmas) are small Gram-negative bacteria that parasitize red blood cells and can cause mild to severe anemia in a wide range of vertebrates, including ruminants. Cattle population in Somalia is around 3.9 million heads, with animals more concentrated around the river areas, mainly in the Juba River and Shabelle River Valleys. Information on hemoplasmas in Sub-Saharan Africa are scarce, mainly in Somalia, where no studies have been performed to date. Accordingly, this study aimed to assess the molecular occurrence of hemoplasmas in 131 cattle blood samples from Somalia. Thirty out of 131 (22.90%; 95% CI: 16.54-30.81%) cattle were infested by ticks: Rhipicephalus pulchellus (68.18%), Amblyomma gemma (18.18%), Amblyomma lepidum (9.09%), Hyalomma marginatum (1.51%), Hyalmomma rufipes (1.51%), and Rhipicephalus pravus (1.51%). A total of 74/131 (56.48%; 95% CI: 47.93-64.67%) cattle were positive for hemotropic Mycoplasma spp. by real-time PCR (qPCR) based on the 16S rRNA gene. Hemoplasma-positive samples were later subjected to species-specific PCR assays for Mycoplasma wenyonii and ‘Candidatus Mycoplasma haematobovis’ based on the 16S rRNA gene. A total of 34/74 (45.94%; 95% CI: 35.07-57.22%) animals were coinfected by both species; 31/74 (41.89%; 95% CI: 31.32-53.26%) and 3/74 (4.05%; 95% CI: 01.39-11.25%) cattle were solely positive to M. wenyonii and ‘Ca. M. haematobovis’, respectively. Six out of 74 (8.1%; 95% CI: 03.77-16.58%) cattle were negative on species-specific conventional PCR assays but tested positive by a semi-nested PCR assay based on the 16S rRNA gene of hemoplasmas. Sequencing of the detected hemotropic Mycoplasma sp. 16S rRNA gene confirmed that animals were infected by M. wenyonii and ‘Ca. M. haematobovis’. To the best of our knowledge, this is the first study on the detection of hemoplasmas in cattle from Somalia.
... Overall, only 20 relevant and accessible studies were identified with the first published in 1975 (Edelsten, 1975) and the most recent in December 2022 (Ferrari et al., 2022). Only two papers on ticks and TBPs were published in the 1980s, and none between 1990 and 2010 (Tables 2, 3). ...
... In terms of parasite and pathogen detection, about 50% of the articles reviewed used molecular techniques while the remaining 50% used methods such as serological analysis, microscopy and bacteriological approaches (Table 3). In both regions of Somalia and Somali region of Ethiopia, only four studies used advanced molecular methodology to detect TBPs (Frangoulidis et al., 2021;Tomassone et al., 2016Tomassone et al., , 2012Ferrari et al., 2022), and these were studies where the analysis was undertaken in laboratories outside of the region, therefore highlighting the lack of modern laboratories in the regions where the samples were collected. ...
Ticks and tick-borne pathogens (TBPs) of domestic animals in Somalia and neighbouring regions of Ethiopia and Kenya are reviewed to identify knowledge gaps in these regions, where unrestricted livestock movements across borders are common. Major scientific databases, such as PubMed, Web of Science, Scopus, CABI, and Google Scholar were searched, to retrieve articles based on papers published between 1960 and March 2023. Thirty-one tick species representing six genera (Rhipicephalus, Hyalomma, Amblyomma, Haemaphysalis, Ornithodoros and Argas) were reported to infest domestic animals, mainly livestock. Overall, the most represented species were Rhipicephalus pulchellus (up to 60% of specimens identified), followed by Hyalomma dromedarii (up to 57%), Hyalomma truncatum (up to 57%), Amblyomma lepidum (up to 21%), Amblyomma variegatum (up to 21%) and Amblyomma gemma (up to 19%), with morphological characterization being the principal method of tick identification. In addition, 18 TBPs, including zoonotic pathogens (e.g., Crimean-Congo haemorrhagic fever virus), were detected, with Babesia spp., Theileria spp., and Rickettsia spp. being the most commonly reported. Half of the pathogens documented were detected using molecular techniques, while the other half were detected by serology and microscopic techniques. Generally, ticks and TBPs in the region are under-studied, particularly, data relating to pet animals and equines is lacking. Further, the infection intensity and herd prevalence of ticks and TBPs is unclear because of insufficient data and poor approaches to quantitative analysis, making it difficult to propose management policies in the region. There is an urgent need, therefore, for more and better studies, particularly those that take a 'One Health' perspective, focusing on the prevalence and socio-economic impact of ticks and TBPs in animals as well as in humans, so that sustainable control strategies against them can be planned.
... Overall, only 20 relevant and accessible studies were identified with the first published in 1975 (Edelsten, 1975) and the most recent in December 2022 (Ferrari et al., 2022). Only two papers on ticks and TBPs were published in the 1980s, and none between 1990 and 2010 (Tables 2, 3). ...
... In terms of parasite and pathogen detection, about 50% of the articles reviewed used molecular techniques while the remaining 50% used methods such as serological analysis, microscopy and bacteriological approaches (Table 3). In both regions of Somalia and Somali region of Ethiopia, only four studies used advanced molecular methodology to detect TBPs (Frangoulidis et al., 2021;Tomassone et al., 2016Tomassone et al., , 2012Ferrari et al., 2022), and these were studies where the analysis was undertaken in laboratories outside of the region, therefore highlighting the lack of modern laboratories in the regions where the samples were collected. ...
Background
Fourteen-years after the last Rift Valley fever (RVF) virus (RVFV) outbreak, Somalia still suffers from preventable transboundary diseases. The tradition of unheated milk consumption and handling of aborted materials poses a public health risk for zoonotic diseases. Limited data are available on RVF and Brucella spp. in Somali people and their animals. Hence, this study has evaluated the occurrence of RVFV and Brucella spp. antibodies in cattle, goats and sheep sera from Afgoye and Jowhar districts of Somalia.
Methods
Serum samples from 609 ruminants (201 cattle, 203 goats and 205 sheep), were serologically screened for RVF by a commercial cELISA, and Brucella species by modified Rose Bengal Plate Test (mRBPT) and a commercial iELISA.
Results
Two out of 609 (0.3 %; 95 %CI: 0.04–1.2 %) ruminants were RVF seropositive, both were female cattle from both districts. Anti- Brucella spp. antibodies were detected in 64/609 (10.5 %; 95 %CI: 8.2–13.2 %) ruminants by mRBPT, which were 39/201 (19.4 %) cattle, 16/203 (7.9 %) goats and 9/205 (4.4 %) sheep. Cattle were 5.2 and 2.8 times more likely to be Brucella -seropositive than sheep (p = 0.000003) and goats (p = 0.001), respectively. When mRBPT-positive samples were tested by iELISA, 29/64 (45.3 %; 95 %CI: 32.8–58.3 %) ruminant sera were positive for Brucella spp. Only 23/39 (58.9 %) cattle sera and 6/16 (37.5 %) goat sera were positive to Brucella spp. by iELISA.
Conclusions
The present study showed the serological evidence of RVF and brucellosis in ruminants from Afgoye and Jowhar districts of Somalia. Considering the negligence of the zoonotic diseases at the human-animal interface in Somali communities, a One Health approach is needed to protect public health.
Background
Serious disease outbreaks in cattle are usually associated with blood pathogens. This study aims to detect blood pathogens namely Theileria species, Anaplasma species, Candidatus Mycoplasma haemobos and Trypanosoma evansi, and determine their phylogenetic relationships and haemato-biochemical abnormalities in naturally infected cattle.
Methods
Molecular analysis was achieved by PCR amplification and sequencing of PCR amplicons of 18SrRNA gene of Theileria species, 16SrRNA genes of Anaplasma and Mycoplasma species, MPSP genes of T. orientalis and T. sinensis, MSP4 gene of A. marginale, 16SrRNA gene of Candidatus Mycoplasma haemobos, and RoTat1.2 VSG gene of Trypanosoma evansi, in sixty-one (61) clinically ill Kedah-Kelantan x Brahman cattle in Pahang, Malaysia.
Results
A total of 44 (72.13%) cattle were infected with more than one blood pathogen. Theileria species was the blood pathogen with the highest molecular detection rate (72.13, 95% CI 59.83–81.81%). Nucleotide blast analyses of all sequences demonstrated high degree of molecular similarity (98–100%) in comparison with their respective reference sequences. Analysis of 18SrRNA gene sequences of Theileria species and 16SrRNA gene sequences of Anaplasma species revealed Theileria sinensis and Anaplasma platys respectively as additional species detected in these cattle. MPSP-PCR analysis was conducted for further confirmation of T. sinensis. The blood picture of eight infected cattle groups revealed poikilocytosis, anisocytosis, rouleaux formation and degenerative left shift. High mean erythrocyte fragility values were common in infected cattle groups. Anaemia of the macrocytic normochromic type and spherocytes were observed in the T. evansi and Anaplasma platys + Theileria sinensis double species co-infected cattle group. Normocytic normochromic anaemia was observed in the T. sinensis infected cattle group. Significant (p < 0.05) increases in serum liver and kidney parameters, total protein, globulin, total and unconjugated bilirubin and decreased albumin values were observed in the T. evansi infected cattle when compared to clinically healthy cattle.
Conclusion
We present the first evidence of Theileria sinensis-associated bovine anaemia (TSABA) in Malaysian cattle. Because of the high occurrence of bovine theileriosis and detection of A. platys, there is an urgent need for appropriate preventive and control measures against these blood pathogens.
This study aimed to elucidate the epidemiological status of hemoplasma infection and investigate the interaction between Theileria orientalis and hemoplasmas in Japanese Black breeding cows raised in the Kyushu and Okinawa regions. Blood samples were collected from 400 cows from 80 different farms in eight prefectures (five samples per farm and 10 farms per prefecture). Mycoplasma wenyonii (Mw), “Candidatus Mycoplasma haemobos” (CMh), and T. orientalis were examined by polymerase chain reaction (PCR) assay using whole blood samples. PCR results showed that 91.5% (366/400) of cows were positive for bovine hemoplasma: 40.3% were infected with Mw only, 9.5% with CMh only, and 41.8% with both species. T. orientalis was detected in 36% (144/400) of cows. The infection rate of T. orientalis was higher in the grazing group (P<0.001) than in the housed group, while the rate of CMh infection was higher (P<0.05) in the housed group than in the grazing group, suggesting that not only the tick but also other arthropod vectors may contribute to hemoplasma transmission. Although the cows with hemoplasma dual infection showed higher (P<0.05) white blood cell counts compared with hemoplasma-negative cows, there was no difference in hematologic parameters related to the anemia between the hemoplasma-positive and -negative animals. This may indicate that Japanese Black cattle could have resistance to the anemia caused by infection with hemoplasma.
African animal trypanosomiasis (AAT) affects the livestock of 12.3 million Somalis and constrains their development and wellbeing. There is missing data on AAT in the country after the civil war of the 1990s. Therefore, this study has aimed to assess the prevalence of Trypanosoma spp. in 614 blood samples from cattle (n = 202), goats (n = 206) and sheep (n = 206) in Afgoye and Jowhar districts, Somalia using parasitological and molecular methods. Twenty-one out of 614 (3.4%; 95% CI: 2.1–5.2%) and 101/614 (16.4%; 95% CI: 13.6–19.6%) ruminants were positive for Trypanosoma spp. by buffy coat technique (BCT) and internal transcribed spacer 1 (ITS1)-polymerase chain reaction (PCR), respectively. Using ITS1-PCR, the highest prevalence was observed in cattle (23.8%; 95% CI: 18.4–30.1%) followed by goats (17.5%; 95% CI: 12.9–23.3%) and sheep (8.3%; 95% CI: 5.1–12.9%). A total of 74/101 (73.3%; 95% CI: 63.5–81.6%) ruminants were shown coinfection with at least two Trypanosome species. The four T. brucei-positive samples have tested negative for T. b. rhodesiense, by the human-serum-resistance-associated-PCR. Trypanosoma evansi, T. godfreyi, T. vivax, T. brucei, T. simiae and T. congolense were the Trypanosoma species found in this study. This is the first study on the molecular detection of Trypanosoma sp. in ruminants in Somalia. Further investigations and control measures are needed to manage Trypanosomiasis spreading in the country. Studies should also focus on the detection of T. b. rhodesiense in the country.
Hemoplasmas (hemotropic mycoplasmas) are small pleomorphic bacteria that parasitize the surface of red blood cells of several mammalian species including cattle, goats, and humans, causing infectious anemia. However, studies on hemoplasmas have been neglected and to date, there are no studies on bovine and caprine hemoplasmas in Uganda or the entire East African region. In this study, a polymerase chain reaction (PCR) assay targeting the 16S rRNA gene was used to investigate the presence of hemoplasma in 409 samples (cattle = 208; goats = 201) collected from Kasese district, western Uganda. Results showed that 32.2% (67/208) of cattle samples and 43.8% (88/201) of goat samples were positive for hemoplasmas. Sequencing analysis identified Candidatus Mycoplasma haemobos and Mycoplasma wenyonii in cattle, while Candidatus Mycoplasma erythrocervae and Mycoplasma ovis were identified in goats. Statistical analysis showed that goats were at a higher risk of infection with hemoplasmas compared with cattle. To the best of our knowledge, this is the first molecular evidence of hemoplasmas in bovine and caprine animals in Uganda and the entire east African region.
We here present annotated lists of names of Candidatus taxa of prokaryotes with ranks between subspecies and class, proposed between the mid-1990s, when the provisional status of Candidatus taxa was first established, and the end of 2018. Where necessary, corrected names are proposed that comply with the current provisions of the International Code of Nomenclature of Prokaryotes and its Orthography appendix. These lists, as well as updated lists of newly published names of Candidatus taxa with additions and corrections to the current lists to be published periodically in the International Journal of Systematic and Evolutionary Microbiology, may serve as the basis for the valid publication of the Candidatus names if and when the current proposals to expand the type material for naming of prokaryotes to also include gene sequences of yet-uncultivated taxa is accepted by the International Committee on Systematics of Prokaryotes.
Tick-borne diseases (TBD) cause enormous losses for farmers. Backyard raising comprises majority of the livestock population in the Philippines, but TBD information in backyard livestock is scarce. In this study, 48 cattle and 114 water buffalo samples from Quezon province, Philippines were molecularly screened for tick-borne pathogens. Anaplasmamarginale (16.67%) and hemoplasma (20.99%) were detected in the samples. A. marginale infection (P=0.0001) was significantly higher in cattle, while hemoplasma infection (P=0.011) was significantly higher in water buffaloes. A. marginale isolates from this study were highly similar to previous isolates from the Philippines while Mycoplasma wenyonii and Candidatus Mycoplasma haemobos were the identified hemoplasma species. Our findings reveal additional information on the TBD situation of Philippine backyard livestock.
Mycoplasma haemocanis is prevalent in the endangered Darwin’s fox (Lycalopex fulvipes) in its main stronghold, Chiloé Island (Chile). The origin of the infection, its dynamics, its presence in other fox populations and the potential consequences for fox health remain unexplored. For 8 years, hemoplasmal DNA was screened and characterized in blood from 82 foxes in Chiloé and two other fox populations and in 250 free-ranging dogs from Chiloé. The prevalence of M. haemocanis in foxes was constant during the study years, and coinfection with “Candidatus Mycoplasma haematoparvum” was confirmed in 30% of the foxes. Both hemoplasma species were detected in the two mainland fox populations and in Chiloé dogs. M. haemocanis was significantly more prevalent and more genetically diverse in foxes than in dogs. Two of the seven M. haemocanis haplotypes identified were shared between these species. Network analyses did not show genetic structure by species (foxes versus dogs), geographic (island versus mainland populations), or temporal (years of study) factors. The probability of infection with M. haemocanis increased with fox age but was not associated with sex, season, or degree of anthropization of individual fox habitats. Some foxes recaptured years apart were infected with the same haplotype in both events, and no hematological alterations were associated with hemoplasma infection, suggesting tolerance to the infection. Altogether, our results indicate that M. haemocanis is enzootic in the Darwin’s fox and that intraspecific transmission is predominant. Nevertheless, such a prevalent pathogen in a threatened species represents a concern that must be considered in conservation actions.
IMPORTANCE Mycoplasma haemocanis is enzootic in Darwin’s foxes. There is a higher M. haemocanis genetic diversity and prevalence in foxes than in sympatric dogs, although haplotypes are shared between the two carnivore species. There is an apparent tolerance of Darwin’s foxes to Mycoplasma haemocanis.
Background:
Camel trypanosomiasis or surra is of great concern in Somalia, since the country possesses the largest one-humped camel (Camelus dromedarius) population in the world. Civil war in Somalia has resulted in the destruction of educational, research, economic and social structures, making the country scores very low for most humanitarian indicators. Previous studies on detection of Trypanosoma species in Somali camels have only been performed during the 1990s using standard trypanosome detection methods (STDM). Considering the lack of state-of-the-art knowledge on camel trypanosomiasis in Somalia, the present study aimed to assess the prevalence of Trypanosoma spp. in three districts of Somalia.
Methods:
A total of 182 blood samples from C. dromedarius from nomadic and dairy farms were evaluated using STDM, serological (CATT/T. evansi) and molecular (ITS1-PCR) methods.
Results:
All samples were negative for Trypanosoma spp. by STDM. A total of 125/182 (68.7%, 95% CI: 61.4-75.3%) camels were seropositive for T. evansi by CATT/T. evansi. Camels reared in nomadic system were more likely to be seropositive for T. evansi than those under dairy production system (OR: 5.6, 95% CI: 2.1-15.2, P = 0.0001). Five out of 182 (2.7%, 95% CI: 0.9-6.3%) camels tested positive for Trypanosoma sp. by ITS1-PCR. Sequencing of the ITS1 region of the Trypanosoma species detected herein revealed that camels were infected with T. evansi and T. simiae.
Conclusions:
Trypanosoma evansi is highly prevalent in camels from the Banadir region of Somalia, particularly in nomadic herds. To our knowledge, this is the first study to confirm infections with T. evansi and T. simiae in Somali camels through DNA sequencing. Our data highlight the need for implementation of adequate control measures aiming to reduce the impact on camel production in the country.
Background: Hemotropic mycoplasmas (aka hemoplasmas) are small bacteria which cause infectious anemia in
several mammalian species including humans. Information on hemoplasma infections in Cuban bovines remains
scarce and no studies applying molecular methods have been performed so far. The aim of the present study was
to utilize real-time PCR and sequence analysis to investigate dairy cattle and buffalo from Cuba for the presence of
bovine hemoplasma species.
Results: A total of 80 blood samples from 39 buffalo and 41 dairy cattle were investigated for the presence of
Mycoplasma wenyonii and “Candidatus Mycoplasma haemobos” using two species-specific real-time TaqMan PCR
assays. PCR results revealed overall 53 (66.2%; 95% CI: 55.3–75.7%) positive animals for M. wenyonii and 33 (41.2%;
95% CI: 31.1–52.2%) for “Ca. M. haemobos”; the latter were all co-infections with M. wenyonii. The sample
prevalences were similar in cattle and buffalo. Based on the sequence analysis of the nearly full-length 16S rRNA
gene from two cattle and two buffalo, the presence of M. wenyonii and “Ca. M. haemobos” was confirmed.
Statistical analysis revealed that buffalo and cattle one year of age or older were more frequently infected with
M. wenyonii or “Ca. M. haemobos” than younger animals. PCR-positivity was not associated with anemia; however,
the infection stage was unknown (acute infection versus chronic carriers).
Conclusions: The high occurrence of bovine hemoplasma infections in buffalo and dairy cattle may have a
significant impact on Cuban livestock production. To the best of our knowledge, this is the first molecular evidence
of bovine hemoplasma species infection in dairy cattle and buffalo from Cuba and the Caribbean.
The African buffalo ( Syncerus caffer ), a mammal species whose population is declining, can play a role as a reservoir or carrier of a wide number of arthropod-borne pathogens. Translocation procedures have been used as an alternative approach for species conservation. However, the veterinary aspects of this sort of procedures are extremely important to minimize the impact on animal health. In order to detect Bartonella and haemoplasmas, two important group of bacterial that have an impact in both human and animal health, EDTA whole-blood samples were screened for the presence of these bacterial pathogens by molecular techniques. As a result, a molecular occurrence of 4.1 and 15.4% for Bartonella spp. and haemoplasmas, respectively, was reported among 97 wild buffaloes sampled during a translocation procedure from Marromeu to Gorongosa Reserve, Mozambique. Additionally, phylogenetic analyses of the obtained sequences were conducted. At least, three bovine-associated pathogens, namely B. bovis , M. wenyonii and ‘ Candidatus M. haemobos’, as well as a probably new Bartonella genotype/species were detected in S. caffer. Further studies are needed in order to determine whether these bacterial species may cause impact in buffaloes and other sympatric ruminant species living in the release site.
Case history and clinical findings:
A dairy cow, from a herd in the Waikato region of New Zealand, was reported with regenerative anaemia on 12 September 2014. Testing of blood from the animal using PCR assays for Theileria orientalis produced a negative result for both Chitose and Ikeda types.
Laboratory findings:
Using PCR and DNA sequencing, blood from the cow was positive for Candidatus Mycoplasma haemobos. Further testing of another 12 animals from the case herd, 27 days after the affected cow was first reported, showed 11 animals were positive for Candidatus M. haemobos or Mycoplasma wenyonii in the PCR. None of these cattle were clinically anaemic or positive for T. orientalis Ikeda type using PCR. A convenience sample of 47 blood samples from cattle throughout New Zealand, submitted to the Investigation and Diagnostic Centre (Ministry for Primary Industries) for surveillance testing for T. orientalis Ikeda, was selected for further testing for bovine haemoplasmas. Of these samples, 6/47 (13%) and 13/47(28%) were positive for M. wenyonii and Candidatus M. haemobos, respectively. There was no difference in the proportion of samples positive for the bovine haemaplasmas between cattle with anaemia that were negative for T. orientalis (6/20, 33%), or without anaemia or T. orientalis (10/18, 56%), or from cattle herds experiencing anaemia and infection with T. orientalis Ikeda type (3/9, 33%).
Diagnosis:
Bovine haemoplasmosis.
Clinical relevance:
The presence of bovine haemoplasmas in blood does not establish causality for anaemia in cattle. Diagnosis of anaemia associated with haemoplasmosis would require exclusion of other causes of regenerative anaemia and an association of the agent with anaemia in affected cattle herds. The data collected in this study did not provide evidence that bovine haemoplasmas were associated with a large number of outbreaks of anaemia in cattle in New Zealand.
The aims of this study were to determine the prevalence of hemoplasmas in a rural Brazilian settlement’s population of human beings, their dogs and horses, highly exposed to tick bites; to identify the tick species parasitizing dogs and horses, and analyze factors associated with their infection. Blood samples from 132 dogs, 16 horses and 100 humans were screened using a pan-hemoplasma SYBR green real-time PCR assay followed by a species-specific TaqMan real-time PCR. A total of 59/132 (44.7%) dog samples were positive for hemoplasmas (21 Mycoplasma haemocanis alone, 12 ‘Candidatus Mycoplasma haematoparvum’ alone and 21 both). Only 1/100 (1.0%) human sample was positive by qPCR SYBR green, with no successful amplification of 16S rRNA or 23 rRNA genes despite multiple attempts. All horse samples were negative. Dogs >1 year of age were more likely to be positive for hemoplasmas (p = 0.0014). In conclusion, although canine hemoplasma infection was highly prevalent, cross-species hemoplasma transmission was not observed, and therefore may not frequently occur despite overexposure of agents and vectors.
Inference of multiple sequence alignments (MSAs) is a critical part of phylogenetic and comparative genomics studies. However, from the same set of sequences different MSAs are often inferred, depending on the methodologies used and the assumed parameters. Much effort has recently been devoted to improving the ability to identify unreliable alignment regions. Detecting such unreliable regions was previously shown to be important for downstream analyses relying on MSAs, such as the detection of positive selection. Here we developed GUIDANCE2, a new integrative methodology that accounts for: (i) uncertainty in the process of indel formation, (ii) uncertainty in the assumed guide tree and (iii) co-optimal solutions in the pairwise alignments, used as building blocks in progressive alignment algorithms. We compared GUIDANCE2 with seven methodologies to detect unreliable MSA regions using extensive simulations and empirical benchmarks. We show that GUIDANCE2 outperforms all previously developed methodologies. Furthermore, GUIDANCE2 also provides a set of alternative MSAs which can be useful for downstream analyses. The novel algorithm is implemented as a web-server, available at: http://guidance.tau.ac.il.
© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.
This study describes a seroepidemiological survey on Rift Valley
fever (RVF) among small ruminants and their close human contacts in
Makkah, Saudi Arabia. A total of 500 small ruminants (126 local, 374
imported) were randomly selected from the sacrifice livestock yards
of Al-Kaakiah slaughterhouse, in the holy city of Makkah, during the pilgrimage season 1432 H (4–9 November 2011). In addition, blood
samples were collected from 100 local workers in close contact with
the animals at the slaughterhouse. An RVF competition multi-species
enzyme-linked immunosorbent assay (ELISA) detecting anti-RVF
virus immunoglobulin G (IgG)/immunoglobulin M (IgM) antibodies
and an RVF IgM-specific ELISA were used for serological
investigations. In total, 84 (16.8%) of the 500 sacrificial sheep and
goats tested seropositive in the competition ELISA but no IgM
antibodies were detected in the IgM-specific assay. All seropositive
samples, comprising 17.91% of the imported animals and 13.49% of
the local ones, were therefore designated positive for anti-RVF virus
IgG antibody. Among the local personnel working in close contact
with the animals, 9% tested seropositive in the RVF competition
ELISA. The study indicates that two factors may increase the
likelihood of an RVF outbreak among sacrificial animals and
pilgrims: i) the large-scale importation of small ruminants into Saudi
Arabia from the Horn of Africa shortly before the pilgrimage season,
and ii) the movement of animals within Saudi Arabia, from the RVFendemic
south-western area (Jizan region) to the Makkah region,
particularly in the few weeks before the pilgrimage season. From
these findings, it is recommended that i) all regulations concerning the
import of animals into Saudi Arabia from Africa should be rigorously
applied, particularly the RVF vaccination of all ruminants destined for
export at least two weeks before exportation, and ii) the movement of
animals from the RVF-endemic south-western area (Jizan region) of
Saudi Arabia to the Makkah region should be strictly prohibited.
A fully illustrated guide to 48 species of ticks of most importance as agents of disease in domestic animals in Africa. It uses a character matrix system, supported by a glossary in which the states of each identification character are illustrated.
Ticks are common on horses, but there is a dearth of contemporary data on infestation prevalence, predominant species, and tick-borne disease agents important in this host. To determine the species of ticks most common on horses and the prevalence of equine exposure to and infection with tick-borne disease agents, ticks and blood samples were collected from 73 horses during May, June, and July of 2010. Adult ticks were identified to species, and antibodies to Ehrlichia spp., Anaplasma spp., and Borrelia burgdorferi were identified using indirect fluorescence antibody assay, a commercial point-of-care enzyme-linked immunosorbent assay, or both. In total, 1,721 ticks were recovered at the majority (85%) of equid examinations. Amblyomma americanum (L.) was the most common tick collected (1,598 out of 1,721; 92.9%) followed by Dermacentor variabilis (Say, 1821) (85 out of 1,721; 4.9%) and Amblyomma maculatum Koch, 1844 (36 out of 1,721; 2.1%); single specimens of Ixodes scapularis Say, 1821 and Dermacentor albipictus (Packard, 1869) were also identified. Antibodies reactive to Ehrlichia spp. were found in 18 out of 73 (24.7%) of horses tested, and were more commonly identified in horses with moderate or high tick infestations than those with low tick infestations (P < 0.001). These data support A. americanum as the most common tick species infesting horses in central Oklahoma from May through July and suggest horses are also commonly exposed to an Ehrlichia sp.
The present study evaluated the effect of hemoplasmosis on cattle productivity. Prevalence of bovine hemoplasma was examined by polymerase chain reaction (PCR) using whole blood samples collected from 93 breeding cows and their 71 calves in Hokkaido, Japan. Monthly milk production records and other clinical data were compared between Mycoplasma wenyonii (Mw)-infected, "Candidatus Mycoplasma haemobos" (CMh)-infected, co-infected and PCR-negative groups. Blood chemical parameters were obtained from the 93 cows and 64 calves. PCR results showed that 89.2% (83/93) of cows and 14.1% (10/71) of calves were positive for bovine hemoplasma. Based on productivity data obtained from the 93 cows, Mw-infected, CMh-infected and co-infected cows had significantly lower monthly milk yield compared to PCR-negative cows. Furthermore, decline in milk yield was prolonged in CMh-infected and co-infected groups. No significant differences were found for other clinical findings among the four groups. Calf birth weight tended to be lower for Mw-infected, CMh-infected and co-infected groups compared to the PCR-negative group. There were no significant differences in all blood parameters of cows and calves among the four groups. In addition, no significant differences were found in any parameter between hemoplasma-infected and PCR-negative calves.
We report a major update of the MAFFT multiple sequence alignment program. This version has several new features, including
options for adding unaligned sequences into an existing alignment, adjustment of direction in nucleotide alignment, constrained
alignment and parallel processing, which were implemented after the previous major update. This report shows actual examples
to explain how these features work, alone and in combination. Some examples incorrectly aligned by MAFFT are also shown to
clarify its limitations. We discuss how to avoid misalignments, and our ongoing efforts to overcome such limitations.
Bovine hemoplasmas are bacteria found on the erythrocyte surface or free in the plasma of cattle. The aim of the present study was to evaluate the occurrence of 'Candidatus Mycoplasma haemobos' ('C. M. haemobos') in Holstein and Jersey cattle raised in Londrina and surroundings, northern region of the State of Parana, Southern Brazil. PCR testing directed to 16S rRNA gene fragment was performed to investigate the occurrence and characterize the molecular identity of 'C. M. haemobos'. A total of 264/433 (60.97%) blood samples were positive by PCR. Further alignment of 500-bp amplicons to available sequences at the GenBank database showed high identity (100%) to 'C. M. haemobos'. To the author's knowledge, this is the first molecular confirmation of the hemoplasma 'C. M. haemobos' in cattle from Brazil. Moreover, 'C. M. haemobos' was observed in high occurrence in dairy cattle, and may have significant impact in livestock production.
We developed a real-time PCR procedure followed by melting curve analysis using the green fluorescence dye SYBR Green I for rapid detection and differentiation of hemplasmas in cattle. Analysis of the melting temperature (Tm) of the PCR products allowed for differentiation of the 2 bovine hemoplasmas, Mycoplasma wenyonii and a provisional species, `Candidatus Mycoplasma haemobos' (a synonym of `Candidatus M. haemobovis'). The Tm (mean ± S.E.) of the PCR products from the bovine hemoplasmas were 86.98 ± 0.12°C for M. wenyonii and 82.04 ± 0.27°C for `Candidatus M. haemobos' in the melting experiments. The protocol described in the present study can decrease the time to results by simultaneous detection and differentiation of the two hemoplasmas in cattle. By using this protocol, we examined hemoplasma prevalence in 109 cattle in Miyagi Prefecture and found that 67 (61.5%) were infected with M. wenyonii, 25 (22.9%) were infected with `Candidatus M. haemobos' and 14 (12.8%) were infected with both.
Concomitantly with an outbreak of fatal anaplasmosis in a cattle herd in Switzerland in 2002, we detected two bovine hemoplasma
species in diseased animals: Mycoplasma wenyonii (formerly Eperythrozoon wenyonii) and a second, novel bovine hemoplasma species later designated “Candidatus Mycoplasma haemobos” (synonym, “Candidatus Mycoplasma haemobovis”). The second species was characterized by a shorter 16S rRNA gene. The aims of the present study were
to provide a detailed molecular characterization of this species, to develop specific quantitative real-time PCR assays for
the two bovine hemoplasma species, and to apply these assays in order to evaluate the prevalence and clinical significance
of the hemoplasmas. Sequencing of the near-complete 16S rRNA gene of the second hemoplasma revealed that it was 94% identical
to that of Mycoplasma haemofelis, an anemia-inducing feline hemoplasma species, but less than 85% identical to that of the bovine hemoplasma M. wenyonii. Using the newly developed assays, a total of 159 animals from the anaplasmosis outbreak were reexamined. In addition, we
tested 57 clinically ill and 61 healthy Swiss cattle, as well as 47 calves. Both hemoplasmas were highly prevalent in adult
cattle but occurred rarely in calves. Animals from the herd with the fatal anemia outbreak were more frequently infected with
M. wenyonii and exhibited higher M. wenyonii blood loads than animals with unrelated diseases and healthy animals. Coinfections may increase the pathogenicity and clinical
significance of bovine hemoplasmosis.
Hemotropic mycoplasmas (hemoplasmas) are the causative agents of infectious anemia in several mammalian species. Their zoonotic
potential has recently been substantiated by the identification of a feline hemoplasma isolate in an immunocompromised human
patient. Although species-specific diagnostic molecular methods have been developed, their application as screening tools
is limited due to the species diversity of hemoplasmas. The goals of this study were to develop a universal hemoplasma screening
assay with broad specificity based on the SYBR green PCR principle, to compare the assay with hemoplasma-specific TaqMan PCR,
and to analyze potential tick vectors and human blood samples to address the zoonotic potential. The newly developed PCR assay
based on the 16S rRNA gene amplified feline, canine, bovine, porcine, camelid, and murine hemoplasmas, as well as Mycoplasma penetrans and Mycoplasma pneumoniae. The lower detection limit for feline and canine hemoplasmas was 1 to 10 copies/PCR. The assay exhibited 98.2% diagnostic
sensitivity and 92.1% diagnostic specificity for feline hemoplasmas. All 1,950 Ixodes ticks were PCR negative, suggesting that Ixodes ticks are not relevant vectors for the above-mentioned hemoplasma species in Switzerland. None of the 414 blood samples derived
from anemic or immunocompromised human patients revealed a clear positive result. The SYBR green PCR assay described here
is a suitable tool to screen for known and so-far-undiscovered hemoplasma species. Positive results should be confirmed by
specific TaqMan PCR or sequencing.
The natural transmission routes of the three feline haemotropic mycoplasmas--Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum', and 'Candidatus Mycoplasma turicensis' (CMt)--are largely unknown. Since CMt has been detected in the saliva of infected cats using PCR, we hypothesised that direct transmission via social or aggressive contact may occur. The aim of this study was to evaluate this transmission route. CMt-positive saliva and blood samples were obtained from three prednisolonetreated specific pathogen-free (SPF) cats that were infected intraperitoneally with CMt. Five SPF cats were inoculated with CMt-positive saliva or blood subcutaneously to mimic cat bites, and five cats were inoculated orally with blood or oronasally with saliva to mimic social contact. Blood samples were monitored for CMt infection using quantitative real-time PCR and for seroconversion using a novel western blot assay. Neither oronasal nor subcutaneous inoculation with CMt-positive saliva led to CMt infection in the recipient cats, as determined by PCR, independent of prior prednisolone treatment. However, when blood containing the same CMt dose was given subcutaneously, 4 of the 5 cats became PCR-positive, while none of the 5 cats inoculated orally with up to 500 microL of CMt-positive blood became PCR-positive. Subsequently, the latter cats were successfully subcutaneously infected with blood. All 13 CMt-exposed cats seroconverted. In conclusion, CMt transmission by social contact seems less likely than transmission by aggressive interaction. The latter transmission may occur if the recipient cat is exposed to blood from an infected cat.
A new approach to rapid sequence comparison, basic local alignment search tool (BLAST), directly approximates alignments that optimize a measure of local similarity, the maximal segment pair (MSP) score. Recent mathematical results on the stochastic properties of MSP scores allow an analysis of the performance of this method as well as the statistical significance of alignments it generates. The basic algorithm is simple and robust; it can be implemented in a number of ways and applied in a variety of contexts including straightforward DNA and protein sequence database searches, motif searches, gene identification searches, and in the analysis of multiple regions of similarity in long DNA sequences. In addition to its flexibility and tractability to mathematical analysis, BLAST is an order of magnitude faster than existing sequence comparison tools of comparable sensitivity.
Three different species of hemoplasmas have been described in rodents, Mycoplasma coccoides, ‘Candidatus Mycoplasma haemomuris’ and ‘Candidatus Mycoplasma haemosphiggurus’. Additionally, potentially novel hemoplasma species have been detected in wild rodents from Brazil, including capybaras (Hydrochoerus hydrochaeris). Capybaras are the largest rodent in the world and are well adapted to live within close proximity to humans, which increases the risk to spread of zoonotic pathogens. Herein, we investigate the occurrence and genetic diversity of hemoplasmas infecting free-ranging capybaras from southern Brazil. Blood samples and ticks from 17 capybaras were collected. Packed cell volume and total plasma protein were measured, DNA was extracted, and further screened by species-specific and pan-hemoplasma PCR assays targeting the 16S rRNA gene of hemoplasmas. Sixteen out of 17 (94.12%; 95% CI: 73.02–98.95%) were anemic. Only one young female was hypoproteinemic. All capybaras were infested by adults and nymphs of Amblyomma dubitatum ticks. Using the PCR assay targeting the 16S rRNA gene of M. coccoides, 13/17 (76.47%; 95% CI: 52.74–90.44%) capybaras were positive for hemoplasmas. When DNA samples were tested by the pan-hemoplasma PCR, 16/17 (94.12%; 95% CI: 73.02–98.95%) animals were positive. One out of 11 (9.09%) adult ticks salivary glands tested positive for hemoplasma by the pan-hemoplasma PCR assay. Sequencing and phylogenetic analysis of the 16S and 23S rRNA gene fragments confirmed that animals were infected by a novel hemotropic Mycoplasma sp. previously reported in capybaras from Brazil. Additionally, sequencing and phylogenetic analysis of the 23S rRNA gene from three hemoplasma-positive capybaras samples from a previous study performed in midwestern Brazil also confirm our findings. Based on phylogenetic and Neighbor-Net network analysis of the 16S rRNA and 23S rRNA genes, the name ‘Candidatus Mycoplasma haematohydrochoerus’ is proposed for this novel organism.
Globally, hemotropic mycoplasmas (hemoplasmas) comprise an emerging or remerging bacteria group that attaches to red blood cells of several mammal's species and in some cases, causing hemolytic anemia. Herein, we assessed the occurrence, genetic diversity, the factors coupled to mammals infection, and the phylogeographic distribution of hemoplasmas in sylvatic and synanthropic mammals and their associated ectoparasites from Brazil. We collected spleen and/or blood samples from synanthropic rodents (Rattus rattus [N=39] and Mus musculus [N=9]), sylvatic rodents (Hydrochoerus hydrochaeris [N=14]) and opossums (Didelphis albiventris [N=43]). In addition, ticks (Amblyomma spp. [N=270] and lice (Polyplax spinulosa [N=6]) specimens were also sampled. Using a PCR targeting the 16S rRNA region, out of 48 small rodents, 14 capybaras and 43 opossums DNA samples, hemoplasma DNA was found in 25%, 50%, and 32.5% animals, respectively. Besides, we reported hemoplasma DNA in Amblyomma sp. (22.2% [2/9]) and lice (100% [2/2]) pools samples from rats, and one female A. sculptum DNA sample (3% [1/33]) obtained from a capybara. Additionally, and in agreement with ML analysis, the network analyses showed a clear phylogenetic separation among the hemoplasmas genotypes found in the different host species sampled, thus, suggesting the absence of cross-species hemoplasmas transmission between the mammals trapped. Finally, using the NTC network analysis, we reported the same 16S rRNA Mycoplasma genotype circulating in Rattus sampled in Brazil, Hungary, and Japan.
Purpose
Haemoparasitic diseases are among the important factors that threaten cattle health and productivity especially in the sub-Saharan region. In Nigeria, their detection using sensitive molecular techniques is scanty. This study was designed to investigate and to reevaluate the repertoire of haemoparasites of cattle in Ibadan, Nigeria with a comparative evaluation of light microscopy (LM) and polymerase chain reaction (PCR) methods.
Methods
Blood samples from 100 cattle slaughtered at Ibadan abattoirs were examined using LM and PCR techniques for haemoparasite detection. The PCR reactions using three primer sets targeting the 16S rRNA genes for Hemoplasma spp. and Anaplasma/Ehrlichia spp. and 18S rRNA genes of Babesia/Theleiria spp. were done. A few randomly selected amplicons from each set were sequenced and analysed.
Results
A total infection rate of 34% by LM including Hemoplasma spp. (17%), Anaplasma spp. (16%), microfilaria (5%) and Trypanosoma spp. (12%) was recorded. While, 86% positivity was recorded with PCR amplification as follows: Hemoplasma spp. (64%), Babesia/Theleiria spp. (46%) and Anaplasma/Ehrlichia spp. (5%). Comparison of LM and PCR findings showed that no LM Anaplasma spp.-positive samples and 7 out of the 17 LM hemoplasma-positive cattle were confirmed by PCR. In addition, LM led to misdiagnosis of 46 Babesia/Theleiria spp.-positive samples. Amplicon sequencing and phylogenetic analysis of Babesia/Theileria spp.-positive samples revealed Theileria velifera and Theileria annulata. In the Anaplasma/Ehrlichia spp.-positive samples, only Anaplasma marginale was characterized. Mycoplasma wenyonii, “Candidatus Mycoplasma haemobos” and Pseudomonas fluorescens like were characterized among the hemoplasma-infected cattle.
Conclusions
The first report of “Candidatus Mycoplasma haemobos” and Pseudomonas fluorescens like in Nigerian cattle is herewith documented. The alarming LM misdiagnosis of haemoparasites during this study confirms its limitations as it fails to identify many parasites and emphasizes the need for inclusion of molecular techniques to improve their detection. The study also shows for the first time the high prevalence of haemotropic mycoplasma in Nigerian cattle via molecular diagnostic methods, thus indicating a strong need for the investigation of their zoonotic implications.
Hemoplasmosis caused by Mycoplasma spp. have been associated with major economic losses in the global dairy production. Hemoplasma studies in the Philippines are limited despite its potential impact. This study mainly aimed to detect the presence of hemoplasma species in dairy water buffaloes and cattle and know their ectoparasite biodiversity in Bohol, Philippines. Detection of Mycoplasma spp. was performed using peripheral blood smear examination (PBSE) and standard PCR using whole blood samples collected from 100 dairy water buffaloes and 40 dairy cattle. Available records on the average annual, monthly and daily milk production were compared between PCR-positive and PCR-negative animals. Ectoparasites were manually collected and identified. While PBSE results were all negative, PCR testing showed that 80% (80 water buffaloes and 32 cattle) were positive for Mycoplasma spp. On the other hand, a total of 1436 ectoparasites were collected (609 Haematopinus and 827 Rhipicephalus spp.). DNA sequencing revealed that obtained sequences (193 bp) from 7 animals were 99.5 to 100% similar to registered Mycoplasma wenyonii sequences. The study reports the first molecular characterization of M. wenyonii in the Philippines and probably the first detection in dairy water buffaloes in Southeast Asia.
Mycoplasma wenyonii, a hemoplasma infecting cattle, was never detected in France. In 2014, evocative inclusions were observed in erythrocytes from cattle presenting milk drops, anemia, and edema in Brittany (France). A survey was then initiated to investigate the epidemiological situation and correlate mycoplasma detection with clinical signs. For this purpose, a new PCR assay targeting polC gene was designed. Comparative results with published PCR assays place this new one as more specific, allowing a one-step diagnosis without further sequencing. A total of 181 cows were included in this study and 4.97% (n = 9) were positive, resulting in the first molecular identification of M. wenyonii in France. All positive animals presented anemia, edema and milk drop. When selecting animals presenting evocative clinical signs, the prevalence of M. wenyonii in Brittany was estimated to 25.6%. Further studies are needed to evaluate the importance of the infection, the implication of arthropods and the existence of asymptomatic carriers.
The way that some parasites and pathogens persist in the hostile environment of their host for long periods remains to be resolved. Here longitudinal field surveys were combined with laboratory experiments to investigate the routes of transmission and infection dynamics of such a pathogen—a wild rodent haemotropic bacterium, specifically a Mycoplasma haemomuris‐like bacterium. Flea‐borne transmission, direct rodent‐to‐rodent transmission, and vertical transmission from fleas or rodents to their offspring were experimentally quantified, and indications were found that the main route of bacterial transmission is direct, although its rate of successful transmission is low (~20%). The bacterium's temporal dynamics was then compared in the field to that observed under a controlled infection experiment in field‐infected and lab‐infected rodents, and indications were found, under all conditions, that the bacterium reached its peak infection level after 25–45 days and then decreased to low bacterial loads, which persist for the rodent's lifetime. These findings suggest that the bacterium relies on persistency with low bacterial loads for long‐term coexistence with its rodent host, having both conceptual and applied implications.
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Hemotrophic mycoplasmas (HMs) are associated with anemia and other disease complexes in a wide range of livestock and wild animals. Two bovine HM species have been identified to date, i.e. Mycoplasma wenyonii and ‘Candidatus Mycoplasma haemobos’. The study aim was to develop quantitative real-time PCR assays (qPCRs) to detect and quantify M. wenyonii and ‘C. M. haemobos’ and to apply these assays to DNA samples extracted from bovine blood collected in Germany (n = 220) from 22 herds. The qPCR assays specific for M. wenyonii and ‘C. M. haemobos’ were designed using the gapN of the respective hemoplasma species as gene target which encodes the NADP-dependent glyceraldehyde 3-phosphate dehydrogenases (GAPN). The sensitivity of both assays was 10 genome equivalents per reaction, corresponding to 2500 genome equivalents per ml blood. No cross-reactivity with non-target bovine HMs. and other bovine pathogens was observed. Bovine HM DNA was detected in 137 samples (62.27%) with 118 samples (53.64%) being positive for ‘C.M. haemobos’ and 19 samples (8.64%) being positive for M. wenyonii. Thereof, 11 animals (5.00%) were co-infected with both bovine HM species. The found herd prevalence for `C. M. haemobos` was 100.00%, and for M. wenyonii 36.36% with mean bacterial loads of 3.7 × 10⁷ `C. M. haemobos`/mL blood and of 4.29 × 10⁵M. wenyonii/mL blood respectively. Clinical and economic relevance of bovine HM species should be goal of future studies for which the novel gapN qPCR assays can serve as a valuable diagnostic tool.
Two species of hemotropic mycoplasmas (HM) are known to infect large domestic ruminants, Mycoplasma wenyonii and ‘Candidatus Mycoplasma haemobos’. Although HM has been described in cattle worldwide, data in water buffaloes (Bubalus bubalis) remain scarce. Accordingly, the aim was to determine the occurrence of HM in water buffaloes from northeastern Brazil. A total of 101/290 (34.83%) buffaloes were positive for HM (16 M. wenyonii alone, 6 ‘Ca. M. haemobos’ alone and 79 both). This was the first report of M. wenyonii infection in ruminants from Brazil. Clinical signs of hemoplasmosis in buffaloes remain unknown.
There has been considerable debate as to whether global risk from vector-borne diseases will be impacted by climate change. This has focussed on important mosquito-borne diseases that are transmitted by the vectors from infected to uninfected humans. However, this debate has mostly ignored the biological diversity of vectors and vector-borne diseases. Here, we review how climate and climate change may impact those most divergent of arthropod disease vector groups: multivoltine insects and hard-bodied (ixodid) ticks. We contrast features of the life cycles and behaviour of these arthropods, and how weather, climate, and climate change may have very different impacts on the spatiotemporal occurrence and abundance of vectors, and the pathogens they transmit.
The 16S rRNA gene of Eperythrozoon (Haemobartonella) wenyonii, a wall-less hemotrophic prokaryote currently classified as a rickettsia, was sequenced to determine the relationship of this organism to other wall-less prokaryotes. Comparison to the GenBank data base showed that this hemotrophic organism is a Mycoplasma (family Mollicutes). Phylogenetic analysis of 16S rRNA genes indicated that this and other recently sequenced 16S rRNA genes of hemotrophic bacteria form a new, separate branch which shares a node in common with the pneumoniae group of mycoplasmas. This result will require that Eperythrozoon wenyonii be reclassified as a Mycoplasma. A main point of this study is that this and related hemotrophic bacteria represent an entirely new group of pathogens among the mycoplasmas.
The 16S rRNA gene of Eperythrozoon (Haemobartonella) wenyonii, a wall-less hemotrophic prokaryote currently classified as a rickettsia, was sequenced to determine the relationship of this organism to other wall-less prokaryotes. Comparison to the GenBank data base showed that this hemotrophic organism is a Mycoplasma (family Mollicutes). Phylogenetic analysis of 16S rRNA genes indicated that this and other recently sequenced 16S rRNA genes of hemotrophic bacteria form a new, separate branch which shares a node in common with the pneumoniae group of mycoplasmas. This result will require that Eperythrozoon wenyonii be reclassified as a Mycoplasma. A main point of this study is that this and related hemotrophic bacteria represent an entirely new group of pathogens among the mycoplasmas.
The present study was carried out in a herd with concurrent infections of Mycoplasma wenyonii and 'Candidatus M. haemobos', to investigate if transplacental and/or vector-borne transmission is possible for one or both bovine haemoplasma species. For this purpose blood samples were collected from 38 mother animals and their newborn calves; as well as from 17 uninseminated cows twice three months apart. In addition, 311 mosquitoes and blood-sucking flies (Diptera: Culicidae, Tabanidae, Muscidae) were cought near the animals. DNA was extracted from all samples, followed by real-time PCR analysis. In 10.5% of neonate calves, that were born to cows harbouring both haemoplasmas, M. wenyonii and/or 'Candidatus M. haemobos' positivity was detected. Copy numbers in positive samples from cows and their calves indicated that - in comparison with M. wenyonii - 'Candidatus M. haemobos'-bacteraemia had usually lower levels. In samples of uninseminated cows the rate of infection with the latter species decreased. These findings may explain why M. wenyonii was significantly more frequently detected in blood-sucking flies, than 'Candidatus M. haemobos'. In conclusion, molecular evidence is provided for the first time on the transplacental transmission of bovine haemoplasmas. Regarding their spread by blood-sucking arthropods, new potential vectors were identified, i.e. the horn fly (Haematobia irritans), the stable fly (Stomoxys calcitrans) and two species of horse flies (Tabanus bovinus, T. bromius).
"Candidatus Mycoplasma haemobos" is a hemoplasma species found in cattle and has been recently reported in Switzerland and Japan. In this study, "Candidatus Mycoplasma haemobos" was shown to occur in cattle and buffalo in tropical China by PCR amplification and sequence analysis of the 16S rRNA gene from blood samples. Based on the 16S rDNA sequence, a specific PCR assay was developed. Occurrence of "Candidatus Mycoplasma haemobos" in cattle and buffalo in Guangxi, China, was determined by examining 25 buffalo blood samples, 12 yellow cattle blood samples and 42 dairy cow blood samples. The results showed that 32% (8/25) of buffalo, 41.7% (5/12) of yellow cattle, and 14.3% (6/42) of dairy cows were positive for "Candidatus Mycoplasma haemobos", respectively. Direct sequencing of representative PCR products confirmed that the amplified partial 16S rDNA sequence represented "Candidatus Mycoplasma haemobos". This is the first report of "Candidatus Mycoplasma haemobos" in buffalo, yellow cattle, and dairy cows in China.
Haemotrophic mycoplasmas are unculturable eperythrocytic pathogens that are found in a wide range of domestic and wild animals. In this study an outbreak of haemotrophic mycoplasmosis in cattle herds in Northern Germany is reported. Affected animals exhibited anaemia and depression and infection was confirmed following microscopic examination of blood smears and on PCR. Sequence analysis indicated that in addition to infection with Mycoplasma wenyonii, animals were infected with a novel bovine haemotrophic mycoplasma Candidatus Mycoplasma haemobos.
The ticks Hyalomma (Euhyalomma) impeltatum Schulze & Schlottke, 1930 and H. (E.) somalicum Tonelli Rondelli, 1935 [a species resurrected for “Hyalomma ? species” of Hoogstraal (1956) and H. erythraeum of Kaiser & Hoogstraal (1968)] are tentatively considered to belong to the H. (E.) asiaticum group of closely related species. Amongst other features that are fairly similar, males of H. impeltatum can be distinguished from those of H. somalicum by the oval posterior margin of the conscutum, a narrow, subtriangular parma, the lack of ventral sclerotised plaques on median, paramedian and 4th festoons, and an incomplete to complete ivory-coloured stripe on the dorsal aspect of the leg segments; whereas males of H. somalicum have a broad but only slightly convex posterior conscutal margin, in most cases no parma, well-developed sclerotised ventral plaques on all festoons, and only a small ivory-coloured spot on the dorsal aspect of the leg segments. Females of H.
impeltatum can be distinguished from those of H. somalicum by the bulging rather than flat preatrial fold of the genital aperture. All parasitic stages of both ticks are illustrated and redescribed, and the characteristics that distinguish the adults from those of other closely related species are detailed. Larger domestic and wild ungulates are the principal hosts of the adults of both ticks. Nymphs and larvae of H. impeltatum parasitise rodents, leporids, birds and lizards, whereas the hosts of the immature stages of H. somalicum are unknown. H.
impeltatum is widely distributed in Africa north of the equator, Arabia, the Near East and south-western part of Central Asia; in contrast, H. somalicum has a more limited distribution in East Africa and possibly the Arabian Peninsular. Data on their possible disease relationships are also provided.
Currently, a lack of consensus exists on how best to perform and interpret quantitative real-time PCR (qPCR) experiments. The problem is exacerbated by a lack of sufficient experimental detail in many publications, which impedes a reader's ability to evaluate critically the quality of the results presented or to repeat the experiments.
The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines target the reliability of results to help ensure the integrity of the scientific literature, promote consistency between laboratories, and increase experimental transparency. MIQE is a set of guidelines that describe the minimum information necessary for evaluating qPCR experiments. Included is a checklist to accompany the initial submission of a manuscript to the publisher. By providing all relevant experimental conditions and assay characteristics, reviewers can assess the validity of the protocols used. Full disclosure of all reagents, sequences, and analysis methods is necessary to enable other investigators to reproduce results. MIQE details should be published either in abbreviated form or as an online supplement.
Following these guidelines will encourage better experimental practice, allowing more reliable and unequivocal interpretation of qPCR results.
A new method for determining nucleotide sequences in DNA is described. It is similar to the "plus and minus" method [Sanger, F. & Coulson, A. R. (1975) J. Mol. Biol. 94, 441-448] but makes use of the 2',3'-dideoxy and arabinonucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase. The technique has been applied to the DNA of bacteriophage varphiX174 and is more rapid and more accurate than either the plus or the minus method.
This paper records, for the first time in New Zealand, the occurrence of Eperythrozoon wenyoni in the blood of a cow. Inoculation of infected blood into two splenectomized calves resulted in a heavy parasitaemia with a moderate anaemia in one calf The other calf remained refractory to infection. The possible importance of E. wenyoni is discussed.
Approximately 10 of 100 young heifers that had recently delivered their first calf--members of a large Colorado dairy herd--had a syndrome of swollen teats and distal portions of the hind limbs, prefemoral lymphadenopathy, transient fever, rough coat, decreased milk production, and subsequent weight loss and reproductive inefficiency. Acute clinical signs of disease were associated with large numbers of Eperythrozoon wenyonii seen on blood smears, and resolution of signs correlated with reduction or disappearance of the parasite. Other known causes of peripheral edema could not be documented. The parasite was transmitted to 4 of 7 nonlactating dairy cows destined to be culled and a splenectomized calf via IV inoculation of blood from parasitemic heifers, but clinical signs of infection were not induced.