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Babesia caballi, Theileria equi and several species of rickettsias are agents of vector-borne diseases that affect equines. The objective of this study was to detect infections by B. caballi and T. equi in horses and to identify rickettsias in horses and ticks in the western region of Cuba. Two nPCR assays were developed and standardized for the detection of B. caballi and T. equi. Blood samples from horses and ticks were collected. Identification by blood smear and molecular detection and identification of B. caballi, T. equi and Rickettsia spp. were carried out. Intraerythrocytic formations compatible with B. caballi and T. equi were observed. The nPCR showed that 25 % of the samples were positive for B. caballi, 73 % for T. equi and 20 % showed coinfection. The results were confirmed with the partial sequencing of the genes bc48 (B. caballi) and ema-1 (T. equi). The sequencing of the 18S rRNA gene of T. equi demonstrated the presence of at least two genotypes of T. equi isolates in Cuba. The real time qPCR assay and sequencing revealed the presence of Rickettsia amblyommatis in A. mixtum and Rickettsia felis in D. nitens. Conclusions: These results constitute the first piece of molecular evidence of B. caballi and T. equi in horses and the first report of R. felis in D. nitens in Cuba, which broadens the knowledge about the distribution of pathogens and the spectrum of potential vectors contributing to the strengthening of management and control programs. [ABSTRACT FROM AUTHOR]

Vector-borne diseases in horses are an emerging and re-emerging problem worldwide, because they have a huge economic impact on affected equine herds. Equine piroplasmosis is considered the most important vector-borne disease affecting the equine species, which includes horses, donkeys, mules and zebras. This disease is caused by the intra-erythrocyte parasites B. caballi and T. equi, mainly in tropical, subtropical and temperate climate regions, where the disease is endemic. These hemoparasites are responsible for significant economic losses in the equine industry, including costs for veterinary treatment, abortions, decreased performance, and death of animals in acute cases of the disease; besides the imposition of international restrictions on the export or participation in equestrian sporting and exhibition events of animals seropositive to B. caballi or T. equi. The cELISA assays for T. equi and B. caballi are the diagnostic tests recommended by the World Organization for Animal Health for the international movement of equines. Different treatment therapies are described for the control and sterilization of both hemoparasites in infected equines. This paper aimed at providing information on the main aspects related to equine piroplasmosis. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The intraerythrocytic parasites Babesia caballi and Theileria equi are the etiological agents of equine piroplasmosis. This disease affects different horse breeds and it has a significant economic impact on the horse industry. The objective of the present study was to determine the presence of B. caballi and T. equi by blood smear and their association with the hematological parameters. Fifty-eight blood samples were taken from clinically healthy horses, which were diagnosed with B. caballi and T. equi infections by blood smear; in addition, the hematological examination of the red and white series was carried out. The mean value of all the hematological parameters evaluated was among the physiological values described for the horse species. From the total samples analyzed, intraerythrocytic formations compatible with B. caballi and T. equi were observed in 3/58 (5.2 %) and 8/58 (13.8 %) animals, respectively. Of these animals, 6/11 (10.3 %) had hematocrit values lower than those physiological. The association between positive animals and possible risk factors (age, sex, presence of ticks and contact with bovines) was evaluated. The statistical analysis showed significant differences when evaluating the relationship between hematocrit values and positive samples to T. equi. [ABSTRACT FROM AUTHOR]

Molecular markers are valuable tools for researches on the genetic diversity, population and evolutionary structure of important infectious agents, and they have a great impact on the design and implementation of control strategies. For the typing of pathogenic microorganisms, several methods have been developed differing in the discriminative power, reproducibility and easy of interpretation. The multi-locus sequence typing was proposed in 1998 as a universal and definitive method for the characterization of bacteria, using the pathogen Neisseria meningitidis as the object of study. There is currently an increasing number of multilocus sequence typing protocols developed and used in epidemiological researches at different scales, as well as in biology and structure studies of different microbial populations, pathogenicity analysis and bacterial evolution. In the present work, the main typing schemes based on this methodology for the study of hemoparasites that affect cattle. [ABSTRACT FROM AUTHOR]

The bovine anaplasmosis is a disease of cattle caused by the rickettsia Anaplasma marginale. The major surface proteins encoded by genes with conserved sequences are potential candidates for vaccines. Among these proteins, MSP1a is expressed by a single copy gene (msp1a). The molecular weight of MSP1a varies among A. marginale isolates from different geographical regions due to the presence, in a variable number, of a sequence of 28-29 amino acids in tandem repeats. In the present study, the sequences of the repeats and of the microsatellite of the protein MSPla of two isolates of A. marginale from bovines in a farm of the Cuban western region were examined. Both isolates belongs to the genotype G, and the isolate 21G presented the repeats T-B-B-C and the 25G of the types T-C. The analysis of these repeated sequences are very important because they include markers for transmissibility by ticks and are of great value for the future development of a vaccine candidate against this hemoparasite. [ABSTRACT FROM AUTHOR]

Anaplasma marginale is a vector-borne rickettsia with high incidence in tropical and subtropical regions that infects the mature erythrocytes of ruminants. Recent studies with proteomic and genomic approaches have identified more than 20 immunogenic proteins within the outer membrane portion, in addition to the major surface proteins MSP1-MSP5 previously characterized. The aim of this study was to verify that the genes encoding for the proteins VirB9-1, VirB9-2, VirB10 and Ef-Tu of A. marginale Habana strain are transcribed during infection in cattle. The RNA was purified from blood of an experimentally infected animal with A.marginale Habana strain. After reverse transcription, virB9-1, virB9-2, virB10 and EF-Tu genes were amplified by PCR and transcripts of all genes analyzed were detected. [ABSTRACT FROM AUTHOR]

The bacterial isolates SUR - 6, DiP/2012 and Oct/Y were isolated from silkworms (Bombyx mori L.) infected naturally, which presented evident symptoms of bacterial flacheria. The goal of this study was to identify the bacterial pathogens associated with symptoms observed in silkworm breeding areas of Cuba, to establish control and sanitation measures. The morphological, biochemical and physiologic study allowed to identify these isolates as Enterobacter cloacae (SUR - 6), Serratia marcescens (DiP /2012), and Acinetobacter baumanii (Oct/Y), which are informed widely in the literature like pathogens of insects. [ABSTRACT FROM AUTHOR]

Recently, a gammaherpesvirus was described in domestic cats (FcaGHV1). The goal of the present study was to investigate the presence of FcaGHV1 in Swiss domestic cats and analyze potential risk factors. Blood samples from 881 cats presented to veterinarians in all Swiss cantons and from 91 stray cats and neoplastic tissue samples from 17 cats with lymphoma were evaluated. FcaGHV1 was detected by real-time PCR targeting the glycoprotein B gene, followed by sequencing. Blood samples were also tested for feline hemoplasmas, feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV). The molecular prevalence of FcaGHV1 was 6.0% (95% confidence interval (CI), 4.5–7.8%) in cats presented to veterinarians and 5.5% (95% CI, 1.8–12.4%) in stray cats. FcaGHV1 PCR-positive cats originated from 19/26 Swiss cantons. Factors significantly associated with FcaGHV1 detection included male sex, age >3 years, nonpedigree status and co-infection with FIV and hemoplasmas. Moreover, FeLV viremia tended to be associated with FcaGHV1 detection. High FcaGHV1 blood loads were found more frequently in FeLV-viremic cats and less frequently in hemoplasma-infected cats than in uninfected cats. Clinical information was unavailable for most of the 881 cats, but leukemia, carcinoma and cardiomyopathy were reported in FcaGHV1-positive cats. None of the tissue samples from the 17 cats with lymphoma tested positive for FcaGHV1. Sequence analyses revealed homogeneity among the Swiss isolates and >99.7% identity to published FcaGHV1 sequences. In conclusion, FcaGHV1 is present in Switzerland with a similar prevalence in cats presented to veterinarians and in stray cats. The pathogenic potential of FcaGHV1 needs further evaluation. [ABSTRACT FROM AUTHOR]

Dogs can be affected by several species of the genus Babesia causing important diseases in different regions of the world. Among these species are Babesia canis, B. vogeli, B. rossi, B. gibsoni, and B. vulves. The aim of this study was to detect the presence of Babesia spp. in stray dogs from Havana province. Sixty dogs were studied and subjected to haematological examination, blood smear and PCR, which amplified 18S rRNA. Three animals (5 %) showed hematocrit values higher than reference values while 23 (38.3 %) showed lower hematocrit values, indicating the presence of an anemic syndrome in those dogs. According to the leukogram analysis, five dogs (8.3 %) had leukocytosis and 7 (11.6 %) had leukopenia. Thirty-one animals were infested with Rhipicephalus sanguineus sensu lato ticks and 10 were co-infested with Rhipicepahalus microplus ticks. Twenty percent of the samples tested positive by PCR, while microscopic examination revealed Babesia spp. merozoites in only 13.3 % of the samples. A relationship was observed between PCR-positive animals and the decrease in hematocrit value. There was no association between the presence of ticks and sex in animals positive to Babesia spp. The present study demonstrates the presence of Babesia spp. in stray dogs from Havana province. [ABSTRACT FROM AUTHOR]

Researches on the detection of mycoplasmas in horses are relatively scarce. However, there are reports about their association to the respiratory-reproductive complex. In Cuba, the presence of these microorganisms in horses is unknown, so the objective of this work was to detect the presence of microorganisms of the class Mollicutes associated to the respiratory-reproductive complex in horses from Mayabeque, Cuba complex in horses from San José de las Lajas, Mayabeque. For this purpose, 97 animals from four localities of this municipality were analyzed. One hundred and thirty-one exudates (97 nasal and 34 preputial) were collected for diagnosis by microbiological culture and PCR. The presence of Mollicutes was detected in 37.11 % of the samples; of these 88.8 % corresponded to nasal exudates and 11.2 % to preputial exudates. The presence of Mollicutes was detected in all localities. Locality "D" and animals older than two years old had the highest number of positive results. An isolate was obtained from preputial exudate, which was identified as Acholeplasma spp. by biochemical tests. Studies related to the presence of Mollicutes in horses are being carried out for the first time in Cuba, so it is necessary to continue studies related to the epidemiology of these pathogens, as part of the control of the respiratory-reproductive complex in this species. [ABSTRACT FROM AUTHOR]

From the sequencing of the entire genome of Anaplasma marginale isolates, genes encoding membrane proteins of this hemoparasite have been identified, as well as novel immunogens as potential vaccine candidates. However, little is known about the genetic variability of genes encoding membrane proteins of Anaplasma marginale isolates. The aim of this study was to determine the degree of conservation of the genes encoding the VirB9, VirB10 proteins, conjugal transfer protein (CTP) and the elongation factor ( Ef- Tu) in the nucleotide sequence of a Cuban Anaplasma marginale isolate and from the deduced amino acid sequence and compare them with those described for Florida and Saint Maries Anaplasma marginale isolates and Anaplasma (centrale) marginale from Israel . The gene and deduced amino acid sequences showed high degree of identity with the sequences described for other isolates of Anaplasma marginale. [ABSTRACT FROM AUTHOR]

Ticks and the vast array of pathogens they transmit, including bacteria, viruses, protozoa, and helminths, constitute a growing burden for human and animal health worldwide. In Cuba, the major tropical island in the Caribbean, ticks are an important cause of vector-borne diseases affecting livestock production, pet animal health and, to a lesser extent, human health. The higher number of tick species in the country belong to the Argasidae family and, probably less known, is the presence of an autochthonous tick species in the island, Ixodes capromydis . Herein, we provide a comprehensive review of the ticks and tick-borne pathogens (TBPs) affecting animal and human health in Cuba. The review covers research results including ecophysiology of ticks, the epidemiology of TBPs, and the diagnostic tools used currently in the country for the surveillance of TBPs. We also introduce the programs implemented in the country for tick control and the biotechnology research applied to the development of anti-tick vaccines., Competing Interests: The authors declare no conflict of interest.