Potential tick vectors for Theileria equi in Israel.

Theileria equi Mehlhorn and Schein, 1998 (Piroplasmida: Babesiidae) is an important tick-borne pathogen of horses that is highly endemic in many parts of the world, including Israel. The present study evaluated the potential roles of five hard tick species [Hyalomma excavatum Koch, 1844; Hyalomma marginatum Koch, 1844; Rhipicephalus turanicus Pomerantsev 1936; Rhipicephalus annulatus Say, 1821; Haemaphysalis parva (Neumann, 1897) (all: Ixodida: Ixodidae)], previously found to infest horses in Israel, in acting as vectors for piroplasmosis. For this, DNA was extracted from whole ticks and, when possible, from the salivary glands in each species (n = 10-59). Polymerase chain reaction amplification and sequencing of the 18S rRNA gene were used to detect T. equi in 48 of the 127 ticks (37.8%) and in 21 of the 90 extracted salivary glands (23.3%) in all five species. All but two sequences were classified as T. equi genotype A; the remaining two were classified as genotype D. The findings of this study point to Ha. parva and R. annulatus as potential novel vectors of T. equi, and suggest that parasite genotype selection occurs within the tick vector.

Comparison of three acute colic pain scales: Reliability, validity and usability.

A valid, reliable and usable scale is needed for assessing severity of acute abdominal pain in horses. The study aim was to compare three different scales: (1) the equine acute abdominal pain scale (EAAPS); (2) a scale described by Mair and Smith (2005; M and S); and (3) a numerical rating scale (NRS). Forty brief films of horses (35 of colic cases and five of control horses) were randomly presented to 46 equine veterinarians from different countries. Participants, randomly divided into three groups, each used one scale. Five randomly selected films were shown twice for determining intra-observer reliability. Speed, ease of use and face validity of the scales were evaluated based on expert opinion. Response rate was excellent: 89% for the EAAPS (16/18), and 100% for the M and S (18/18) and NRS groups (10/10). The intraclass correlation (ICC) of 0.86 [95% confidence interval (CI); 0.80-0.92] for EAAPS indicated significantly better inter-observer reliability compared to 0.68 for the M and S and 0.71 for the NRS. Moreover, intra-observer reliability of EAAPS (weighted κ 0.95 [95%CI; 0.92-0.98]) was superior to the other scales (weighted κ 0.78, 0.77, for the M and S and NRS, respectively). Other validity measures (convergent, extreme group, predictive validities), usability (time taken to score the films-speed) and the ease of use of the scales were not significantly different. Face validity (endorsement by experts) was better for the M and S scale than for the EAAPS. The EAAPS showed superior reliability, the M and S scale better face validity, with comparable usability and other tests of validity.

Post-partum concentrations of serum progesterone, oestradiol and prolactin in Arabian mares demonstrating normal maternal behaviour and Arabian mares demonstrating foal rejection behaviour.

The aim of this prospective study was to investigate possible endocrine components to foal rejection behaviour in post-partum Arabian mares. Arabian mares were divided into two groups based on their maternal behaviour: (1) mares with normal post-parturient behaviour (8 mares); and (2) mares that demonstrated foal-rejecting behaviour post-partum (15 mares). Most mares were visited and sampled twice, in the first and third days post-partum. Serum samples were used for measurement of progesterone, oestradiol and prolactin concentrations. There were no statistically significant differences in oestradiol, progesterone or prolactin concentrations between the groups. In the rejecting mares, there was a statistically significant decrease in the progesterone (mean±standard deviation, SD, 3.14±6.2ng/dL on day 1 and 0.49±0.18ng/dL on day 3) and prolactin (mean±SD 216.2±325.4ng/mL on day 1 and 145.2±311.4ng/mL on day 3) concentrations between days 1 and 3, while the oestradiol concentration did not change significantly. In the non-rejecting mares, progesterone concentrations decreased significantly (mean±SD 0.8±0.23ng/dL on day 1 and 0.43±0.22 on day 3) while the oestradiol and prolactin concentrations did not change significantly. The oestradiol to progesterone ratio was significantly higher in non-rejecting mares on day 1 (mean±SD 114.8±140.2 on day 1 and 143.4±72.6 on day 3) and this ratio increased significantly from days 1 to 3 in the rejecting mares (mean±SD 47.3±21.1 on day 1 and 122.1±123.7 on day 3).

Analysis of the Association of Climate, Weather and Herd Immunity with the Spread of Equine Encephalosis Virus in Horses in Israel.

It is claimed that the distribution of Culicoides-borne viruses is highly influenced by climate. Equine encephalosis virus (EEV) is a Culicoides-borne orbivirus which affects horses and was recently found to be endemic in Israel. To test whether climate is a crucial factor in the geographical distribution of EEV, we collected blood samples from horses in Israel during the years 2002, 2007 and 2010 and tested them for the abundance of antibodies to EEV. Samples were also collected in 2011 from horses that were seronegative to the virus in 2010, to determine the rate of infection with EEV. It was found that seroprevalence fluctuated between the years and that in each year it was highest in a different climatic region. Interestingly, analysis of infection rate at the different farms showed a negative association with seroprevalence at prior observations. In addition, analysis of precipitation preceding the outbreak of EEV which occurred during 2008 revealed that an extremely dry period existed several months prior to the febrile outbreak with the average precipitation of spring 2008 being significantly lower than the average spring precipitation of the years 1997-2009. It is therefore conjectured that exposure to EEV is not climate specific. Rather, it is highly influenced by herd immunity and weather fluctuations which might change annually. This finding may have important implications for the prediction of the abundance of Culicoides-borne viruses in endemic regions.

Streptococcus equi subspecies equi in horses in Israel: seroprevalence and strain types.

The purpose of this cross-sectional study was to determine the seroprevalence of Streptococcus equi in Israel, to monitor seropositive horses over time and to identify archived strains that were recovered from Israeli horses. A serological survey of 200 healthy horses on 20 farms throughout Israel was performed to detect recent exposure to S equi antigens A and C via indirect ELISA. Seroprevalence was 9.5 per cent (19/200) and positive horses were found in 30 per cent (6/20) of the farms. Sixteen horses that returned a positive serology result were retested three and six months later. Most (12/16) positive horses remained positive, which suggests the presence of animals with persistent infection. Molecular characterisation of S equi strains by sequencing of the SeM gene of 16 archived isolates of S equi that were recovered from clinical cases of strangles between 2008 and 2012 identified two strains: SeM-2 and SeM-28.

Surgical treatment for acute small intestinal obstruction caused by Parascaris equorum infection in 15 horses (2002-2011).

REASONS FOR PERFORMING STUDY: There is a consensus in the veterinary literature that Parascaris equorum (ascarid) impaction carries a poor prognosis. Hence surgery is often delayed and foals are often subjected to euthanasia after diagnosis of ascarid impaction is established. Our clinical impression was that ascarid impaction carries a better prognosis than previously reported. OBJECTIVES: Our expectation was that manually evacuating the impaction into the caecum, thus refraining from opening the small intestine, would improve the prognosis for survival of horses with ascarid impaction. The aim of this study was to examine medical records of horses treated surgically for ascarid impaction, record their clinical findings and evaluate the association of outcome with the method of relieving the impaction. METHODS: The medical records of all horses presented to our hospital between October 2002 and December 2011 that underwent exploratory celiotomy for ascarid impaction were reviewed. Information retrieved from the medical record included surgical findings, surgical technique, complications, short- and long-term survival. The association between categorical variables was assessed using the Fisher's exact test. A P value < 0.05 was considered statistically significant. RESULTS: Fifteen horses fulfilled the inclusion criteria. Eighty percent of horses (12/15) survived to discharge and 60% (6/10) survived for at least one year. Five horses are doing well but are still in convalescence. Horses that underwent small intestinal enterotomy or resection and anastomosis (n = 5) were less likely to survive to discharge (P = 0.022). CONCLUSIONS: In our study, manual evacuation of ascarid impaction into the caecum, while refraining from opening the small intestine resulted in significantly improved survival in horses with ascarid impaction. POTENTIAL RELEVANCE: Refraining from opening the small intestine may be the key to improving surgical outcome in horses with ascarid impaction.

Initial investigation of mortality rates and prognostic indicators in horses with colic in Israel: a retrospective study.

REASONS FOR PERFORMING STUDY: Initial assessment of the mortality rates and prognostic indicators in horses with colic presented to a referral hospital in Israel. OBJECTIVES: To determine mortality rates and to identify potential prognostic indicators in horses undergoing treatment for colic. METHODS: The medical records of 208 colic cases were reviewed and mortality rates calculated including 95% confidence intervals. Mortality rates in surgical cases were calculated separately for strangulating and nonstrangulating lesions as well as for lesions of the large and small intestines. Potential prognostic indicators were identified and evaluated by Student's t test or chi2 test, where appropriate. Those found to be significant (P < 0.05) were evaluated in 2 logistic regression models; one including all horses with colic and one for surgical cases only. RESULTS: The overall mortality rate was 51/208 (25%); 5/72 (7%) in medically treated cases, 46/136 (34%) in surgical cases, 30/50 (60%) in strangulating lesions and 15/85 (18%) in nonstrangulating lesions, 17/27 (63%) in cases involving small intestinal lesions and 28/108 (26%) in cases with large intestinal lesions. Clinical parameters found to be significantly associated with death by univariate analysis were medical/surgical treatment, location of lesion, severity of lesion, mucous membrane colour (MM), capillary refill time (CRT) and heart rate. Using a multivariate logistic regression model, including all cases, medical/surgical treatment, CRT and MM were found to be prognostic indicators and when using the surgical cases alone, only CRT and lesion severity remained related to mortality. CONCLUSIONS: Mortality rates were similar or better than those previously reported in most cases, however, studies from the USA and the UK published better success rates for small intestinal surgeries. Cultural attitudes toward euthanasia may be associated with mortality rates. POTENTIAL RELEVANCE: These results assist clinicians in providing an immediate prognosis based on clinical findings at presentation and contribute to an international database that may aid future research in improving treatment of colic.

Surgical management of sand colic impactions in horses: a retrospective study of 41 cases.

OBJECTIVE: A retrospective review of the medical records of 41 horses requiring abdominal surgery for sand colic. RESULTS: The diagnosis of sand colic was made when sand was found to be the cause of impaction of the gastrointestinal tract during surgical exploration. The most common clinical signs at presentation were abdominal pain, abdominal distension and diarrhoea. A statistically significant association was found between the respiratory rate on arrival and short-term survival. Sand impaction at multiple locations was detected in one-third of the horses. Concurrent pathology was detected in half of the horses. Four horses were euthanased during surgery; of those that recovered from surgery, 35/37 (95%) were discharged from hospital. Short- and long-term complications were similar to those previously reported. Long-term (1 year) survival of the horses discharged was 100%. CONCLUSION: The good prognosis for horses undergoing surgery for the treatment of sand impaction supports early surgical intervention in cases where large amounts of sand are suspected.

Pharmacokinetics of tramadol in horses after intravenous, intramuscular and oral administration.

Tramadol is a centrally acting analgesic drug that has been used clinically for the last two decades to treat moderate to moderately severe pain in humans. The present study investigated tramadol administration in horses by intravenous, intramuscular, oral as immediate-release and oral as sustained-release dosage-form routes. Seven horses were used in a four-way crossover study design in which racemic tramadol was administered at 2 mg/kg by each route of administration. Altogether, 23 blood samples were collected between 0 and 2880 min. The concentration of tramadol and its M1 metabolite were determined in the obtained plasma samples by use of an LC/MS/MS method and were used for pharmacokinetic calculations. Tramadol clearance, apparent volume of distribution at steady-state, mean residence time (MRT) and half-life after intravenous administration were 26+/-3 mL/min/kg, 2.17+/-0.52 L/kg, 83+/-10 min, and 82+/-10 min, respectively. The MRT and half-life after intramuscular administration were 155+/-23 and 92+/-14 min. The mean absorption time was 72+/-22 min and the bioavailability 111+/-39%. Tramadol was poorly absorbed after oral administration and only 3% of the administered dose was found in systemic circulation. The fate of the tramadol M1 metabolite was also investigated. M1 appeared to be a minor metabolite in horses, which could hardly be detected in plasma samples. The poor bioavailability after oral administration and the short half-life of tramadol may restrict its usefulness in clinical applications.

Cattle immune response to botulinum type D toxoid: results of a vaccination study.

Cattle botulism is a food-borne intoxication caused by the ingestion of preformed botulinum neurotoxins (BoNT) of serotypes B, C, or D. Protection in cattle against botulinum intoxication is based on the presence of specific serum neutralizing antibodies upon exposure. Outbreaks in vaccinated cattle have raised concerns about vaccine quality and efficacy. To this end, three different immunization protocols and the effect of maternal anti-BoNT/D antibodies, at the priming dose, were analyzed in 2-month-old dairy calves. Based on previously determined protective anti-BoNT/D antibody levels analyzed in field outbreaks, the immune response to type D toxoids was analyzed using an in-house ELISA system. Here we show that using the current vaccination strategy of using a priming dose in 2-month-old calves followed by booster doses after 4 weeks and annually thereafter, did not result in continuous protective levels of anti-BoNT/D antibodies. As a result of this vaccination protocol, only 15-31% of cattle in parities 1-3 were protected at the time of the annual booster. Vaccination study in calves indicated that adding a 6-month booster dose to the current protocol resulted in continuous protective levels of anti-BoNT/D antibodies well above the cut-off protective levels. The presence of maternally derived anti-BoNT/D antibodies did not interfere with the immune response to toxoids that can be administered to 2-month-old calves.

Seroprevalence of antibotulinum neurotoxin type C antibodies in horses in Israel.

REASONS FOR PERFORMING STUDY: Clostridium botulinum type C is prevalent in Israel and outbreaks recorded in many species, other than horses. Association between levels of anti-BoNT/C antibodies and equine grass sickness (EGS) have been demonstrated but seroprevalence of anti-BoNT/C antibodies in horses has not been reported nor has EGS been reported in Israel. OBJECTIVES: To determine the seroprevalence of specific anti-BoNT/C antibodies in horses in Israel and to determine whether age, breed and gender, or geographical region of farms are potential risk factors for exposure to BoNT/C. HYPOTHESIS: Anti-BoNT/C antibodies are prevalent among horses in Israel and farm and horse-level variables are associated with increased risk for exposure. METHODS: Serum samples from 198 horses were collected and the levels of specific anti-BoNT/C antibodies were determined using enzyme-linked immunosorbent assay (ELISA). For each categorical variable indicator variables were created and the odds ratio (OR) and 95% confidence intervals (95% CI) for the outcome variable were calculated using a univariable and multivariable logistic regression models. RESULTS: A total of 61 (30.8%) horses were ELISA positive for anti-BoNT/C IgG antibodies. The farm and its geographical region were associated significantly with seropositivity, horse-level variables, such as gender and breed, were also associated with seropositivity. Quarter Horse and Warmblood mares placed in the southern region of Israel had the highest odds to be tested positive for anti-BoNT/C IgG antibodies. CONCLUSIONS AND POTENTIAL RELEVANCE: Several farm and various horse-level risk factors for exposure to BoNT/C, found in this study, could be correlated to previously reported risk factors of EGS. Studies are required to determine the predisposing factors that cause EGS, which is apparently not present in Israel.

Quantitative analysis of levels of serum immunoglobulin G against botulinum neurotoxin type D and association with protection in natural outbreaks of cattle botulism.

The recent outbreaks of cattle botulism in vaccinated Israeli dairy cattle prompted us to determine vaccine efficacy and reasons for vaccine failure. Analysis of clinical signs, feeding practice, vaccination history, and epidemic curves enabled us to define a study population in two outbreaks, where high doses of Clostridium botulinum neurotoxin type D (BoNT/D) were evenly consumed by the affected animal groups. Attack rates among unvaccinated 6- to 24-month-old heifers were 96% (55/57) and 85% (53/62). The attack rates in vaccinated parity 1, 2, and >or=3 cows were 40.4% (21/52), 14.3% (4/28), and 5.6% (3/54), respectively. Vaccine efficacies for these cow groups were 52.5%, 83.2%, and 93.4%, respectively. In younger, unvaccinated 2- to 6-month-old calves, presumably protected by maternal antibodies, the attack rate was 24% (17/71). These differences correlated with significant differences in levels of specific anti-BoNT/D antibody in serum by an enzyme-linked immunosorbent assay (ELISA). The ELISA performance for predicting protection was analyzed by receiver operating characteristic analysis and was found to be highly significant, with an area under the curve of 0.941 (standard error, 0.034; 95% confidence interval, 0.875 to 1.008; P < 0.000). No animals with serum ELISA unit levels above 0.33 were affected in these exposed groups. At this cutoff level, the specificity of the ELISA was 100%, sensitivity was 67%, and accuracy was 92%. We concluded that botulinum toxoids can confer adequate protection against natural exposure to lethal doses of BoNT/D; however, the vaccination protocols should be optimized. Our in-house ELISA system will enable us to optimize vaccination protocols in the animal population.

Low seroprevalence of antibodies to Neospora caninum in wild canids in Israel.

The role of domestic dogs in the epidemiology of Neospora caninum as well as the relationship between N. caninum infection of farm dogs and cattle were demonstrated, however, evidence is scarce regarding the role of wild canids in domestic animal neosporosis. The present study was undertaken to evaluate the role of wild canids in the epidemiology of bovine neosporosis in Israel by analyzing the prevalence of antibodies to N. caninum in wild canids. Sera samples were collected from 114 free ranging wild golden jackals (Canis aureus), 24 red foxes (Vulpes vulpes) and nine wolves (Canis lupus), which were collected in Israel during the years 1999-2004. Of a total of 147 wild canids tested antibodies to N. caninum were only found in two golden jackals with IFAT titers of 1:50, and in one red fox and one wolf with IFAT titer of 1:400. The low seroprevalence found in this study (2.7%) indicated that wild canids probably do not have an important role in the epidemiology of N. caninum in Israel. However, since the diet of different species of wild canids and even diverse populations of the same canid species vary, it is possible that other results might be obtained from specific wild canids populations, which scavenge in the vicinity of infected bovines.

Epidemiological study of Culicoides hypersensitivity in horses in Israel.

The variations with breed, gender, age and coat colour in the prevalence of Culicoides hypersensitivity were studied in 408 horses on 18 farms in Israel. Data were gathered by means of questionnaires and the diagnoses were confirmed by direct physical examinations. The prevalence of Culicoides hypersensitivity was 28 per cent. The disease was rare on farms more than 800 m above sea level but was more prevalent at lower altitudes. The most important factors affecting the prevalence of Culicoides hypersensitivity were the farm, breed and age, but gender and colour were not significantly correlated with its prevalence.

Pharmacokinetics of gentamicin C1, C1a and C2 in horses after single intravenous dose.

Gentamicin pharmacokinetics has not been studied in horses. Pharmacokinetics of gentamicin C1, C1a and C2 components following i.v. administration of total gentamicin at 6.6 mg/kg bwt to 6 healthy mature horses was determined. Significant differences in clearance, half-life (t 1/2), and mean residence time (MRT) between the gentamicin Cia and the 2 other components were found. The total body clearance (CL) of gentamicin C1a was 1.62 +/- 0.50 ml/min x kg and similar to the glomerular filtration rate (GFR) reported for horses. The CL of gentamicin C1 and C2 were 1.03 +/- 0.08 ml/min x kg and 1.10 +/- 0.15 ml/min x kg, respectively, and significantly slower than that of gentamicin C1a. The values of apparent volume of distribution at steady state were 0.22 +/- 0.05, 0.26 +/- 0.12 and 0.23 +/- 0.05 l/kg for gentamicin C1, C1a and C2, respectively. The MRT values were mean +/- s.d. 3.6 +/- 0.5, 2.7 +/- 0.3 and 3.5 +/- 0.4 h and the t 1/2 values were 3.1 (2.5-4.0), 2.4 (2.0-3.2) and 33 (2.4-4.3) h (harmonic mean and range) for gentamicin C1, C1a and C2, respectively. The MRT and t 1/2 values for gentamicin C1a were significantly shorter than those of gentamicin C1 and C2. It was concluded that the difference in pharmacokinetics between the gentamicin components has potential pharmacological and toxicological implications.

Clinical signs of West Nile virus encephalomyelitis in horses during the outbreak in Israel in 2000.

Between August and October 2000, 76 horses were reported by veterinary practitioners as having signs of a neurological disorder, varying from an involvement of the spinal cord alone to the entire central nervous system; 15 of the horses died or were euthanased as a result of their grave prognosis or secondary complications. At the same time, an outbreak of West Nile virus infection affected people and birds, principally domestic geese. West Nile virus was isolated from four of the horses with encephalomyelitis and five other horses seroconverted, indicating that the virus was the probable cause of the outbreak in horses. Three of the cases from which the virus was isolated are described briefly and one case is described in detail. This horse behaved abnormally and had general proprioceptive deficits in all four limbs. Its neurological condition deteriorated after two days and severe inspiratory dyspnoea due to a failure to abduct the arytenoids necessitated a tracheostomy. It died on the fourth day and histological lesions were observed in the brain stem and grey matter of the spinal cord.