Trypanosoma evansi Assessment in Equines: A Study in One Decade in an Endemic Area of the Canary Islands,Spain

Trypanosoma evansi was diagnosed in a camel in the Canary Islands for the first time in 1997, and the parasite is still present in a little area of the Archipelago. Since then, the disease has exclusively affected camel herds, and clinical evidence of the infection has not been demonstrated in any other animal hosts. In the frame of a control plan of the disease in the Archipelago, 864 equines located in the infected area were examined during one decade (2001-2010), of which a total of 26 (3%) were serologically positive but showed negative results at parasitological and molecular examinations. FreeCalc, an epidemiological tool to detect presence/absence of disease, showed that the population would be diseased at a confidence level of 99.916%. These results must suppose to implement the control plan against the disease including the equine population present in the surrounding area of the infected camel farms.     

Immunization with Trichophyton verrucosum Vaccine in Hunter/Jumper and Dressage Horses with Naturally Occurring Trichophyton equinum Infection: A Prospective,Randomized, Double-Blinded,Placebo-Controlled Clinical Trial

The aim of the current study was to investigate the efficacy of Trichophyton verrucosum vaccine in hunter/jumper and dressage horses with trichophytosis caused by Trichophyton equinum (cross immunity). A total of 25 hunter/jumper and dressage horses between the ages of 2 and 14 years, naturally infected with Trichophyton equinum, were enrolled and randomly assigned to either a placebo or an intramuscular lyophilized Trichophyton verrucosum vaccine group. Clinical evaluations were done by the same investigator, who scored clinical healing at the beginning, during, and at the end of the treatment and was blinded to allocation to the groups. At the end of the trial, vaccine treatment significantly decreased the investigator's clinical scores (P < .001), and no significant changes were detected in the placebo treatment group. In all of the horses vaccinated, the lesions improved gradually within 7 to 12 days after vaccination. Complete clinical remission was detected within 28 to 42 days, and all treated horses became culture negative within 25 days of starting treatment. No clinical healing was observed in nine untreated control horses throughout the study. No recurrence was observed in treated horses within 10 months after therapy. Results of the current study indicate that inactivated Trichophyton verrucosum vaccine may be a safe and effective therapy for horses with Trichophyton equinum infection.     

Passive transfer of Theileria equi antibodies to neonate foals of immune tolerant mares

Equine babesiosis, a tick transmitted haemoprotozoan disease caused by Theileria equi is globally distributed and responsible for heavy economic losses to the equine husbandry. Equids reared in endemic areas usually pick up infection at an early age and become immune tolerant throughout their life span. We studied the level of passively transferred antibodies in neonate foals born from pre-immuned mares. Latently T. equi infected pre-immuned pony and donkey mares (three each) were selected and T. equi antibody titres in neonates was monitored till 90 days post foaling (DPF) by applying Dot-ELISA on sequentially collected serum samples from foals and their dams. A very high antibody titre was observed in pre-immuned pony and donkey mares. The maximum antibody of 1:60 to 1:80 was observed in pony's and donkey's foal on 2–16 and 2–10 DPF, respectively and thereafter it declined to less than 1:20 on 63–77 and 56–63 DPF. Simultaneously parasite carrying status in neonate foals and their dam was also monitored by applying PCR on blood samples. We could demonstrate PCR amplification in dam's blood samples while no amplification was recorded in neonate's blood samples. This study indicated that new-born foals were born naïve and passively transferred immunity was transitory which wanes after 63–77 DPF.     

A Multiplex Polymerase Chain Reaction Assay for Direct Detection and Differentiation of β-Hemolytic Streptococci in Clinical Samples from Horses

Streptococcus equi subspecies equi, S equi subspecies zooepidemicus, and S dysgalactiae subspecies equisimilis are β-hemolytic Streptococci, often isolated from horses with respiratory or genital diseases. The aim of this study was (i) defining and validating a multiplex polymerase chain reaction (PCR) protocol for identifying these Streptococci in bacterial cultures and for detecting them directly in equine clinical specimens, and (ii) defining and validating a cheap DNA extraction protocol for clinical specimens. When respiratory and genital samples from symptomatic and asymptomatic horses were tested by bacterial culture and by multiplex PCR, all the 150 samples culture-positive for S equi, S zooepidemicus, or S equisimilis were also positive by PCR. Of 150 culture-negative samples, 143 were negative by PCR. Seven samples were positive by PCR but negative by bacteriology. The multiplex PCR protocol described in this study is proven suitable for a sensitive, specific, and rapid detection and identification of S equi, S zooepidemicus, and S equisimilis in cultured bacterial colonies, as well as in clinical specimens from symptomatic or asymptomatic horses. The inclusion of internal control primers in the PCR protocol excludes false-negative results. A cheap DNA extraction method has been also validated for swabs, tracheal aspirates, bronchoalveolar lavage, and guttural pouches lavage samples.     

Etiologic and epidemiologic analysis of bacterial infectious upper respiratory disease in Thoroughbred horses at the Seoul Race Park

Infectious upper respiratory disease (IURD) of Thoroughbred racehorses has been a frequent problem (29.6% of incidence) at the Seoul Race Park (Korea). Risk factors for IURD include the season with a high transfer rate (summer and fall), the stabling period (≤ 3 months), and age (2 to 3 years old), suggesting that the movement and new environment may have depressed the immune system of the horses and decreased their ability to respond properly to pathogens. The bacterial strains (n = 98) isolated from IURD horses included Pseudomonas spp., Escherichia coli, Staphylococcus spp., Streptococcus equi subsp. equi and zooepidemicus.     

Reemergence of Dourine in Italy: Clinical Cases in Some Positive Horses

In 2011, Trypanosoma equiperdum reemerged in Italy, almost 10 years after its last appearance. A total of eight infected horses have been observed to date. Six horses were affected by natural outbreaks of the disease, whereas two were infected experimentally. The aim of this study was to offer a recent perspective on clinical cases of dourine in Europe. Investigation of the clinical aspects confirmed the three stages reported in the literature: stage 1 (genital lesions), stage 2 (cutaneous signs), and stage 3 (nervous signs). The most common signs in the horses under study were notable weight loss, edematous skin eruptions and oedemas of the abdomen, mammary glands and hind legs. Three animals presented neurological signs (lip ptosis of lower lip and ataxia). Infections were paucisymptomatic or asymptomatic in some animals. Hyperthermia was not reported in infected animals and considerable anemia was observed. High antibody titers did not always correspond to clinical signs. Positive polymerase chain reaction test results of blood or tissue (skin, eye swab) often correspond to an advanced stage of the disease. Dourine is a variable disease; owing to its low prevalence and chronic manifestation, it can be difficult to make a quick diagnosis when facing a Dourine-positive horse.     

Diagnosis and treatment of coenurosis in sheep

Coenurosis is a disease of the central nervous system in sheep, caused by Coenurus cerebralis, the larval stage of Taenia multiceps, a tapeworm, which infests the small intestine of carnivores. In 80-90% of cases, the cyst is located in one cerebral hemisphere, whilst in 5-10% of cases, it is localised in the cerebellum; rarely it involves two sites in the brain of the affected animal. Listeriosis, louping-ill, sarcocystosis and polioencephalomalacia and brain abscessation should be considered when formulating a diagnosis of acute coenurosis. In all cases, it is essential to carefully examine the animal and not simply rely on results of ancillary tests (mainly of cerebrospinal fluid examination), as disorders other than coenurosis can be responsible for changes in the results of these tests. Treatment is based on surgical removal of the coenurus cyst after general anaesthesia of the animal; the approach has a very good success rate, especially after accurate localisation of the lesion. Despite that, many farmers may choose to slaughter those sheep fit for marketing for economic reasons and euthanise those in poor condition.     

一例肉鸡大肠杆菌和粪肠球菌混合感染的诊治

本试验对某发病肉鸡场的病死鸡进行了病原学诊断。通过病原分离、生化试验、动物试验、血清型鉴定、PCR扩增等方法对分离细菌进行了鉴定,最终确定为大肠杆菌和粪肠球菌的混合感染。在此基础上进行了药敏试验,筛选出了大肠杆菌的高敏药物阿米卡星和庆大霉素,而粪肠球菌只对某些氟喹诺酮类药物中度敏感,其余均显示为耐药。将阿米卡星、庆大霉素和恩诺沙星用于发病鸡群的治疗,很快控制了疫情,及时减少了经济损失。     

Piroplasmosis in Donkeys—A Hematological and Serological Study in Central Ethiopia

Working donkeys (n = 395) originating from three different districts of the Oromia region of Ethiopia were examined October 2009 and April 2010 for the prevalence of equine piroplasmosis between. Apart from hematological and serological examination, tick vectors were collected and determined. Intraerythrocytic stages of Theileria equi and Babesia caballi were detected in 12.2% and 1.8% of examined stained blood smears, respectively. The seroprevalence of T. equi and B. caballi was 55.7% and 13.2%, respectively. The majority of active infection coincided with anemia. Only 16 donkeys were infested with four species of hard ticks: Rhipicephalus turanicus, Rhipicephalus evertsi evertsi, Rhipicephalus pulchellus, and Amblyomma variegatum.     

The Prevalence of Antibodies against Borrelia burgdorferi Found in Horses Residing in the Northwestern United States

Lyme disease, a bacterial illness caused by Borrelia burgdorferi, is thought to be most prevalent in the heavily tick-infested areas of the northeastern United States. Serum samples from 196 asymptomatic horses residing in the Pacific northwest were tested for the presence of antibodies to Borrelia burgdorferi, using the canine SNAP 4DX (IDEXX Laboratories, Inc., Maine) and the enzyme-linked immunosorbent assay (ELISA). Positive samples were confirmed by Western blot analysis. The ELISA and Western blot analyses identified 29 of 196 horses that had antibodies for Borrelia burgdorferi, whereas the Canine SNAP 4DX only identified 2 of 196 horses as positive for an antibody titer. These results indicate that 14.8% of horses residing in the northwestern United States have been exposed to B. burgdorferi.     

Lineages of Streptococcus equi ssp. equi in the Irish equine industry

Background

Streptococcus equi ssp. equi is the causative agent of ‘Strangles’ in horses. This is a debilitating condition leading to economic loss, yard closures and cancellation of equestrian events. There are multiple genotypes of S. equi ssp. equi which can cause disease, but to date there has been no systematic study of strains which are prevalent in Ireland. This study identified and classified Streptococcus equi ssp. equi strains isolated from within the Irish equine industry.

Results

Two hundred veterinary isolates were subjected to SLST (single locus sequence typing) based on an internal sequence from the seM gene of Streptococcus equi ssp equi. Of the 171 samples which successfully gave an amplicon, 162 samples (137 Irish and 24 UK strains) gave robust DNA sequence information. Analysis of the sequences allowed division of the isolates into 19 groups, 13 of which contain at least 2 isolates and 6 groups containing single isolates. There were 19 positions where a DNA SNP (single nucleotide polymorphism) occurs, and one 3 bp insertion. All groups had multiple (2–8) SNPs. Of the SNPs 17 would result in an amino acid change in the encoded protein. Interestingly, the single isolate EI8, which has 6 SNPs, has the three base pair insertion which is not seen in any other isolate, this would result in the insertion of an Ile residue at position 62 in that protein sequence. Comparison of the relevant region in the determined sequences with the UK Streptococcus equi seM MLST database showed that Group B (15 isolates) and Group I (2 isolates), as well as the individual isolates EI3 and EI8, are unique to Ireland, and some groups are most likely of UK origin (Groups F and M), but many more probably passed back and forth between the two countries.

Conclusions

The strains occurring in Ireland are not clonal and there is a considerable degree of sequence variation seen in the seM gene. There are two major clades causing infection in Ireland and these strains are also common in the UK.     

Cryptosporidiosis in Horses of Urban Areas of Porto Alegre,Rio Grande do Sul,Southern Brazil

The aim of the present study was to detect the presence of Cryptosporidium spp. oocysts in the feces of horses used in military training and in horseback riding activities. A total of 90 fecal samples were collected from three sites in the city of Porto Alegre, state of Rio Grande do Sul, Brazil as follows: 71 samples were obtained from the Brazilian Army, seven from the hospital accredited with the School of Veterinary Medicine of Universidade Federal do Rio Grande do Sul, and 12 from a facility offering animal rest and training. All samples were collected between the months of January and February, 2008. They were screened microscopically for oocysts using the modified Ziehl–Neelsen staining technique. Oocysts were classified on the basis of their microscopic characteristics. The overall infection rate of horses as determined by microscopic analysis was 27.87% (25/90) and was identified only in horses raised in the Brazilian Army, with a rate of 35.21% (25/71). The morphometric analysis of oocysts revealed a mean size of 4.78 × 4.0 μm and a shape index of 1.16. None of the infected horses appeared to be clinically ill.     

Frequency of Rhodococcus equi in Feces of Mares in Central Kentucky

Rhodococcus equi (R equi) pneumonia is an important cause of disease and death in foals. Feces from mares can contain R equi, including virulent R equi, and thus may act as a source of the bacteria at horse breeding farms. A previous report documented that every mare at a farm in central Kentucky shed virulent R equi in at least one of four fecal samples collected serially during the periparturient period. The objective of this study was to assess the extent to which this high prevalence of fecal shedding could be replicated at other horse breeding farms. The frequency of detection of R equi and virulent R equi in fecal samples was studied among 131 mares from 24 farms in central Kentucky. The proportions of fecal samples from mares containing R equi and virulent R equi were 95% and 76%, respectively. These findings indicate that R equi and virulent R equi may be isolated with high frequency from feces of mares at breeding farms in central Kentucky, and that mares are a source of virulent R equi for the environment of their foals.     

Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosis

Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid method whereby DNA is amplified with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specifically designed primers and a DNA polymerase with strand displacement activity. In this study, we used LAMP primer sets designed from EMA-1 and Bc 48 genes for detection of Theileria equi and Babesia caballi infections, respectively. These primer sets specifically amplified DNA of the respective parasites. Both primer sets amplified T. equi and B. caballi up to 10(-6) dilution of 10-fold serially diluted samples. Furthermore, DNA extracted from blood collected from a horse experimentally infected with T. equi was amplified by a T. equi LAMP primer set from days 2 to 35 post-infection, demonstrating the high sensitivity of these primers. Of 55 samples collected from China, 81.8% and 56.3% were positively detected by LAMP for T. equi and B. caballi infections, respectively. In contrast, 91.8% and 45.9% of the 37 samples collected from South Africa were LAMP positive for T. equi and B. caballi, respectively. These results suggest that LAMP could be a potential diagnostic tool for epidemiological studies of equine piroplasmosis.     

Sequence heterogeneity in the 18S rRNA gene within Theileria equi and Babesia caballi from horses in South Africa

A molecular epidemiological survey of the protozoal parasites that cause equine piroplasmosis was conducted using samples collected from horses and zebra from different geographical locations in South Africa. A total of 488 samples were tested for the presence of Theileria equi and/or Babesia caballi using the reverse line blot hybridization assay. Ten percent of the samples hybridized to the Theileria/Babesia genus-specific probe and not to the B. caballi or T. equi species-specific probes, suggesting the presence of a novel species or genotype. The small subunit of rRNA gene (18S; ∼1600 bp) was amplified and sequenced from 33 of these 488 samples. Sequences were compared with published sequences from the public sequence databases. Twelve distinct T. equi and six B. caballi 18S rRNA sequences were identified. Alignments demonstrated extensive sequence variation in the V4 hypervariable region of the 18S rRNA gene within T. equi. Sequence variation was also found in B. caballi 18S rRNA genes, although there was less variation than observed for T. equi. Phylogenetic analysis based on 18S rRNA gene sequences revealed three T. equi clades and two B. caballi clades in South Africa. The extent of sequence heterogeneity detected within T. equi and B. caballi 18S rRNA genes was unexpected since concerted evolution is thought to maintain homogeneity within repeated gene families, including rRNA genes, in eukaryotes. The findings reported here show that careful examination of variants of the 18S rRNA gene of T. equi and B. caballi is required prior to the development of molecular diagnostic tests to detect these parasites in horses. Species-specific probes must be in designed in regions of the gene that are both conserved within and unique to each species.     

Equine Piroplasmosis

Equine piroplasmosis (EP) is a tick-borne protozoal disease of horses, mules, donkeys, and zebras that is characterized by acute hemolytic anemia. The etiologic agents are two hemoprotozoan parasites, Theileria equi (Laveran, 1901) and Babesia caballi (Nutall and Strickland, 1910) that are transmitted primarily by ixodid ticks. Equine piroplasmosis is found globally where tick vectors are present and is endemic in tropical, subtropical, and some temperate regions. Horses infected with B. equi remain seropositive for life; horses infected with B. caballi are seropositive for several years to life. Economic losses associated with EP are significant and include the cost of treatment, especially in acutely infected horses; abortions; loss of performance; death; and restrictions in meeting international requirements related to exportation or participation in equestrian sporting events. Equine babesiosis–free countries limit the entrance of Babesia-seropositive horses into their countries. In the United States a few sporadic outbreaks have occurred in recent years but have been limited due to implementation of stringent control methods. The cELISA for both T. equi and B. caballi is currently the recommended test for international horse transport. Different therapies for control and sterilization of the parasites are discussed.