Synergistic effects of using sodium hypochlorite (bleach) and desiccation in surface inactivation for Toxocara spp.

Toxocara cati and T. canis are parasitic nematodes found in the intestines of cats and dogs respectively, with a cosmopolitan distribution, and the potential for anthropozoonotic transmission, resulting in human toxocariasis. Spread of Toxocara spp. is primarily through the ingestion of embryonated eggs contaminating surfaces or uncooked food, or through the ingestion of a paratenic host containing a third-stage larva. The Toxocara spp. eggshell is composed of a lipid layer providing a permeability barrier, a chitinous layer providing structural strength, and thin vitelline and uterine layers, which combined create a biologically resistant structure, making the Toxocara spp. egg very hardy, and capable of surviving for years in the natural environment. The use of sodium hypochlorite, household bleach, as a disinfectant for Toxocara spp. eggs has been reported, with results varying from ineffective to limited effectiveness depending on parameters including contact time, concentration, and temperature. Desiccation or humidity levels have also been reported to have an impact on larval development and/or survival of Toxocara spp. eggs. However, to date, after a thorough search of the literature, no relevant publications have been found that evaluated the use of sodium hypochlorite and desiccation in combination. These experiments aim to assess the effects of using a combination of desiccation and 10% bleach solution (0.6% sodium hypochlorite) on fertilized or embryonated eggs of T. cati, T. canis, and T. vitulorum. Results of these experiments highlight the synergistic effects of desiccation and bleach, and demonstrate a relatively simple method for surface inactivation, resulting in a decrease in viability or destruction of T. cati, T. canis and T. vitulorum eggs. Implications for these findings may apply to larger scale elimination of ascarid eggs from both research, veterinary, and farming facilities to mitigate transmission.

Proteomic profiling of membrane vesicles from Mycobacterium avium subsp. paratuberculosis: Navigating towards an insilico design of a multi-epitope vaccine targeting membrane vesicle proteins.

Bacteria typically produce membrane vesicles (MVs) at varying levels depending on the surrounding environments. Gram-negative bacterial outer membrane vesicles (OMVs) have been extensively studied for over 30 years, but MVs from Gram-positive bacteria only recently have been a focus of research. In the present study, we isolated MVs from Mycobacterium avium subsp. paratuberculosis (MAP) and analyzed their protein composition using LC-MS/MS. A total of 316 overlapping proteins from two independent preparations were identified in our study, and topology prediction showed these cargo proteins have different subcellular localization patterns. When MVs were administered to bovine-derived macrophages, significant up-regulation of pro-inflammatory cytokines was observed via qRT-PCR. Proteome functional annotation revealed that many of these proteins are involved in the cellular protein metabolic process, tRNA aminoacylation, and ATP synthesis. Secretory proteins with high antigenicity and adhesion capability were mapped for B-cell and T-cell epitopes. Antigenic, Immunogenic and IFN-γ inducing B-cell, MHC-I, and MHC-II epitopes were stitched together through linkers to form multi-epitope vaccine (MEV) construct against MAP. Strong binding energy was observed during the docking of the 3D structure of the MEV with the bovine TLR2, suggesting that the putative MEV may be a promising vaccine candidate against MAP. However, in vitro and in vivo analysis is required to prove the immunogenic concept of the MEV which we will follow in our future studies. SIGNIFICANCE: Johne's disease is a chronic infection caused by Mycobacterium avium subsp. paratuberculosis that has a potential link to Crohn's disease in humans. The disease is characterized by persistent diarrhea and enteritis, resulting in significant economic losses due to reduced milk yield and premature culling of infected animals. The dairy industry in the United States alone experiences losses of approximately USD 250 million due to Johne's disease. The current vaccine against Johne's disease is limited by several factors, including variable efficacy, limited duration of protection, interference with diagnostic tests, inability to prevent infection, and logistical and cost-related challenges. Nevertheless, a multiepitope vaccine design approach targeting M. avium subsp. paratuberculosis has the potential to overcome these challenges and offer improved protection against Johne's disease.

Decreasing polymethyl methacrylate bone cement concentration extends working and setting times in vitro.

OBJECTIVE: To determine setting and temperature properties of diluted polymethyl methacrylate (PMMA) bone cement in vitro to assess utility for vocal fold augmentation in horses. SAMPLES: 4 dilutions of PMMA equivalent to volumes of 15 mL, 20 mL, 25 mL, and 30 mL PMMA powder (PMMAp) in 10 mL solvent. METHODS: For each volume PMMAp, setting times (tset), peak temperatures (Tmax), and times to peak temperature (tmax) were determined using a temperature data logger in a 4-mL volume of PMMA. Injectability was assessed in vitro by documenting the force required to inject 0.2 mL PMMA through an 18-gauge 3.5-inch spinal needle attached to a 6-mL syringe at 1-minute intervals. Working time (twork) was calculated from a linear regression of injectability. RESULTS: Peak temperatures increased with increasing volume of PMMAp: 56 °C, 86 °C, 99 °C, and 101 °C. Times for tset, twork, and tmax were inversely proportional to PMMA concentrations, resulting in tset of 23, 21, 17, and 14 minutes; twork of 22.75, 12.25, 7, and 4 minutes; and tmax of 28, 24, 19, and 16 minutes, respectively, for 15, 20, 25, and 30 mL PMMAp. Pairwise comparisons for all analyses were significant apart from Tmax for 25 and 30 mL PMMAp (P = .96) and twork for 20 and 25 mL PMMAp (P = .06). CLINICAL RELEVANCE: Decreasing the concentration of PMMA bone cement resulted in longer working times and setting times; however, peak temperatures did not differ between the 2 strongest concentrations. Further research is warranted to quantify diluted PMMA properties for in vivo use for vocal fold augmentation in horses.

Serological and molecular surveillance of West Nile virus in domesticated mammals of peninsular Malaysia.

West Nile virus is a mosquito-borne neurotropic pathogen with a wide host range that constitutes a significant risk to public and animal health. There is limited information on WNV infection in domesticated mammals in Malaysia; however, current reports indicate infections in birds, macaques, bats and pigs from Malaysia. In this study, 203 serum samples from cattle, goats, and horses were tested for the presence of anti-WNV IgG using a competitive enzyme-linked immunosorbent assay (c-ELISA). Additionally, using one-step RT-PCR, nasopharyngeal swabs were analyzed for WNV RNA from all 203 animals in this study. The WNV seroprevalence was 32.53% (27/83) at 95% CI (0.2342-0.4319) in cattle, 48.27% (14/29) at 95% CI (0.3139-0.6557) in goats and 53.84% (49/91) at 95% CI (0.4366-0.6373) in horses. Cross-reactive JEV antibodies were detected in two cattle and 34 horses. None of the cattle or goats tested positive for WNV RT-PCR. Seven horses were positive for WNV RT-PCR, a molecular prevalence of 7.69% (7/91) at 95% CI (0.0353-0.1528). This is the first reported detection of WNV in domesticated mammals of Malaysia, a significant addition to the growing evidence that WNV is being transmitted from vectors to susceptible hosts in Malaysia.

The potential risk associated with foodborne pathogens in a watershed: Escherichia coli O157:H7 in dairy cattle.

We carried out an investigation to determine the occurrence of Escherichia coli O157:H7 in dairy herds in the Delaware County watershed and to identify the factors that play a role in the likelihood of presence of this organism among animals on these farms. The pathogen poses risk of environmental degradation and health to the inhabitants. A total of 2162 fecal samples were collected per rectum from a representative sample of cattle on 27 dairy farms. The samples were investigated for the presence of E. coli O157:H by initially enriching using a bacteriological media and detection of the pathogen using real-time polymerase chain reaction technique. Escherichia coli O157:H7 was detected in 74% of the herds in the target population and in 3.7% of samples collected. An additional 54 animals were identified that were infected with O157 non-H7 strains of E. coli among 15 farms. Several putative risk factors were associated with the detection of the pathogen on the enrolled farms included age, housing calves indoors, group housing for calves, housing calves in the calf barn, presence of dogs on the farm, and housing post-weaned calves in a cow barn or heifer barn versus a greenhouse. In conclusion, E. coli O157:H7 was present on the dairy farms of Delaware County and may pose a threat to the people that live and work there. The risk associated with the detection of this pathogen could be reduced by adjusting management factors that were identified in this study.

Circulation of West Nile virus in mosquitoes approximate to the migratory bird stopover in West Coast Malaysia.

Being a tropical country with a conducive environment for mosquitoes, mosquito-borne illnesses such as dengue, chikungunya, lymphatic filariasis, malaria, and Japanese encephalitis are prevalent in Malaysia. Recent studies reported asymptomatic infection of West Nile virus (WNV) in animals and humans, but none of the studies included mosquitoes, except for one report made half a century ago. Considering the scarcity of information, our study sampled mosquitoes near migratory bird stopover wetland areas of West Coast Malaysia located in the Kuala Gula Bird Sanctuary and Kapar Energy Venture, during the southward migration period in October 2017 and September 2018. Our previous publication reported that migratory birds were positive for WNV antibody and RNA. Using a nested RT-PCR analysis, WNV RNA was detected in 35 (12.8%) out of 285 mosquito pools consisting of 2,635 mosquitoes, most of which were Culex spp. (species). Sanger sequencing and phylogenetic analysis revealed that the sequences grouped within lineage 2 and shared 90.12%-97.01% similarity with sequences found locally as well as those from Africa, Germany, Romania, Italy, and Israel. Evidence of WNV in the mosquitoes substantiates the need for continued surveillance of WNV in Malaysia.

A Prospective Study of Growth Rate, Disease Incidence, and Mortality in Kittens Less than 9 Weeks of Age in Shelter and Foster Care.

A prospective study was designed to (a) investigate the rate of daily weight gain among kittens less than 9 weeks old presented to an animal shelter, (b) identify factors (e.g., sex, clinical signs of disease, diet, and medical treatment) that affect daily weight gain, and (c) investigate the mortality of study kittens. The study of 203 kittens was conducted at a managed admission, no-kill animal shelter in upstate New York, USA, from April 2014 through October 2014. Body weight was measured daily from day of intake to adoption or 12 weeks of age. Fecal score, clinical signs of disease, food type, and medical treatments were recorded daily. Lethargy and being female were significantly associated with lower daily weight gain. Despite the challenges of shelter and foster care, the average daily weight gain for study kitten was higher than that reported in other settings such as catteries and laboratories. Five study kittens (2.5%) died or were euthanized. Daily monitoring systems provide opportunities for interventions, increased live outcomes, and improved welfare for kittens in shelter and foster care.

Serum Phosphorylated Neurofilament Heavy Subunit Levels and its Association with the Risk for Catastrophic Injury in Thoroughbred Racehorses.

Neurofilaments are structural proteins that are concentrated in the body and axons of neurons. Damage to the neurons or axons as a result of trauma or infectious diseases leads to the release of neurofilaments into blood and cerebrospinal fluid (CSF). This case-control study was carried out to compare serum levels of phosphorylated neurofilament heavy chain (pNF-H) between clinically healthy Thoroughbred (TB) horses and TB horses that suffered catastrophic musculoskeletal injuries (cMSI), and to investigate the correlation between putative risk factors and serum concentrations of pNF-H in injured horses. Blood samples were collected from clinically healthy horses and from horses that suffered cMSI. The concentration of pNF-H in serum samples was determined using the Phosphorylated Neurofilament H Sandwich enzyme-linked immunosorbent assay kit. A total of 343 horses were enrolled in the study (148 cases and 195 controls). The median serum concentration of pNF-H for controls was 0.0 ng/ml and for cases was 0.07 ng/ml. No significant difference was observed between the 2 groups in racing. The number of lifetime starts was correlated with serum pNF-H concentration in case horses. The serum concentration of pNF-H was higher in case horses that experienced cMSI while training than while racing. The number of lifetime starts is a proxy measure for several risk factors related to cumulative exercise load during the career of racehorses. Measurement of serum concentrations of pNF-H in TB racehorses does not support the hypothesis that subclinical neurologic injury or conditions are associated with catastrophic injury of TB racehorses.

Comparative Efficacy of Topical Ophthalmic Ganciclovir and Oral Famciclovir in Cats with Experimental Ocular Feline Herpesvirus-1 Epithelial Infection.

Purpose: To determine the comparative efficacy of ganciclovir ophthalmic gel and famciclovir oral tablets in cats with experimentally induced ocular feline herpesvirus-1 (FHV-1) epithelial infection. Methods: A randomized, placebo-controlled trial was performed using 16 nonvaccinated, specific pathogen-free cats with experimental FHV-1 infection induced by topical ocular inoculation. Cats received topical ganciclovir 0.15% ophthalmic gel (1 drop 3 times daily, n = 6 cats), oral famciclovir (90 mg/kg twice daily, n = 6), or topical artificial tear gel (1 drop 3 times daily, n = 4) for 14 days. Cats were monitored after inoculation for 30 days. Ophthalmic examinations were performed every 2 days and ocular disease scores calculated. In vivo confocal microscopy was performed, and corneal leukocyte infiltrates quantified. Ocular samples for FHV-1 quantitative polymerase chain reaction (qPCR) and virus isolation assays were collected every 3 days. Hemograms and serum biochemistry panels were performed at intervals. Results: Clinical ocular disease scores and corneal leukocyte infiltrates were significantly lower in the ganciclovir and famciclovir groups compared with placebo, but no significant differences were detected between the antiviral treatment groups. Ocular viral loads determined by qPCR were significantly lower in the ganciclovir group compared with the placebo group, but there were no significant differences between the other study groups. Hemograms and biochemistry panels were unremarkable. Conclusion: Topical application of ganciclovir gel 3 times daily was well-tolerated and displayed similar efficacy at reducing clinical ocular disease scores and corneal inflammation as twice daily oral famciclovir treatment in cats with experimental ocular FHV-1 infection.

Retrospective evaluation of phacoemulsification and aspiration in 182 eyes: Visual outcomes and CDE-predictive value.

OBJECTIVE: To determine the effect of cumulative dissipated energy (CDE) on complications and visual outcome following cataract surgery in dogs. ANIMALS STUDIED: Records of 182 canine eyes that had undergone cataract surgery. PROCEDURES: Retrospective medical record review was conducted on dogs that underwent unilateral or bilateral elective cataract surgery by a single surgeon. Preoperative variables such as age, cataract duration, and pre-existing conditions were recorded. Intraoperative variables such as intraocular lens (IOL) status, phacoemulsification time, average power, irrigation fluid volume, and CDE were recorded. Postoperative complications and visual status at last follow-up were compared to these variables. RESULTS: Eighty-six percent of dogs were visual in one or both eyes at last follow-up (LFU) examination. Median follow-up time was 186 days postoperatively. Eyes that developed glaucoma had a significantly greater mean CDE (77.73 ± 80.11) than eyes that did not develop glaucoma (49.21 ± 38.93). Eyes that remained visual at the LFU examination had a significantly lower mean CDE (48.31 ± 37.14) than eyes that were blind at LFU (82.05 ± 82.73). Significantly lower mean CDEs were found in diabetic eyes, younger eyes, early-operated eyes (<1 month duration), and eyes with preoperative lens-induced uveitis. CONCLUSIONS: Higher CDE may be associated with an increased risk of postoperative glaucoma and loss of vision in dogs. Cumulative dissipated energy may be a useful prognostic indicator for success of canine cataract surgery.

Levels of Serum Phosphorylated Neurofilament Heavy Subunit in Clinically Healthy Standardbred Horses.

Neurofilaments heavy chain proteins (pNF-H) have been identified as useful serum biomarkers for humans and animals with neurologic conditions, some of which can lead to poor performance, and athletic injuries. However, there are no published reports that describe a reference range for serum pNF-H levels in healthy racehorses. This cross-sectional study was carried out to determine the serum concentration of pNF-H in 1,349 samples collected from 1,291 clinically healthy standardbred (SB) racehorses. Data on age, time of sampling (pre-race or post-race), and finishing position during a race were collected. The concentration of pNF-H in serum samples was determined using an enzyme-linked immunosorbent assay (ELISA). The appropriate statistical techniques were used to determine the median serum concentration of pNF-H in these horses, if the serum concentration of pNF-H changed with age, if there were changes in the serum concentration of pNF-H during a race, and if there was an association between serum concentration of pNF-H, and the finishing position for the horse. The median serum concentration of pNF-H in this group of clinically healthy SB horses was 0.0 ng/mL. The concentration of pNF-H in serum was not associated with the age of the horses in this study as was determined by regression analysis. There was no significant change in the serum concentration of pNF-H before and after a race in paired samples. There was no association of serum concentration of pNF-H and the finishing position of the horses after the race. The data from this study supports use of <0.412 ng/mL as a reference interval for measurement of serum levels of pNF-H in SB racehorses as 95% of the collected samples fell into the range 0.0-0.412 ng/mL.

Bile Acids, Direct Bilirubin and Gamma-glutamyltransferase as Prognostic Indicators for Horses with Liver Disease in the Eastern United States: 82 Cases (1997-2019).

Serum biochemistry results and presence of fibrosis on liver biopsies are frequently used as prognostic indicators in horses with liver dysfunction. The objective of this retrospective multicenter study was to determine if the magnitude of abnormal liver specific biochemical tests such as bile acids (BA), direct bilirubin and gamma-glutamyltransferase (GGT), or the presence of fibrosis reported on liver biopsies was associated with prognosis in horses with liver dysfunction. Eighty-two horses older than one year, examined at four referral hospitals in the eastern United States, with BA values greater than 30 µmol/L and having 6-months or more follow-up were included in the study. The association of the maximal BA, GGT and direct bilirubin values of each horse with survival was determined by logistic regression analysis. The presence or absence of fibrosis (non-quantitated) on a liver biopsy was compared between survivors and non-survivors by chi square test. The degree of increase in BA concentration and GGT activity was not related to outcome (OR 0.9999, 95% CI 0.9923 - 1.007, P = 0.97, and OR 1.0, 95% CI 0.9997 - 1.001, P = 0.31 respectively). Direct bilirubin concentration was positively associated with non-survival (OR 1.95, 95% CI 1.34-3.19, P = 0.0023). The presence of fibrosis was not associated with outcome (P = 0.37). These findings suggest that the magnitude of GGT and BA values or the mere presence of fibrosis on liver histopathology should not be used as prognostic indicators. In this study, direct bilirubin values were a better predictor of outcome.

In vivo confocal microscopic features of naturally acquired canine herpesvirus-1 and feline herpesvirus-1 dendritic and punctate ulcerative keratitis.

OBJECTIVE: To describe the in vivo confocal microscopy (IVCM) features of the corneal epithelium and stroma in dogs and cats with herpetic dendritic ulcerative keratitis. ANIMALS: 6 client-owned dogs and 10 client-owned cats with herpetic dendritic ulcerative keratitis (affected group) and 10 dogs and 10 cats from specific-pathogen-free laboratory colonies (nonaffected group). PROCEDURES: After complete ophthalmic examination, IVCM corneal examination was performed on the clinically diseased eyes of animals in the affected group and on both eyes of animals in the nonaffected group. Results by species were compared between groups. RESULTS: In the affected group, all 6 dogs had unilateral ocular lesions (total, 6 eyes examined), whereas 7 cats had unilateral lesions and 3 cats had bilateral lesions (total, 13 eyes examined). For the nonaffected group, 20 cat eyes and 20 dog eyes were examined. Corneal epithelial morphological abnormalities were identified in all examined eyes of animals in the affected group and in no examined eyes of the nonaffected group. Hyperreflective punctate opacities and inflammatory cells were present in all epithelial layers in examined eyes of affected animals but were absent in nonaffected animals. Similarly, Langerhans cells and anterior stromal dendritic cells were identified in corneas of eyes examined for animals in the affected group but not in any eye of animals in the nonaffected group. Stromal changes were less consistent in the affected group, but absent in the nonaffected group. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that herpetic dendritic ulcerative keratitis in dogs and cats is associated with microanatomic corneal abnormalities that can be detected by IVCM.

Attrition of Thoroughbred and Standardbred racehorses at New York Racetracks due to exercise and non-exercise related fatalities during the 2016 - 2019 racing seasons.

A longitudinal retrospective study was carried out to investigate the attrition of Thoroughbred (TB) and Standardbred (SB) horses racing at New York racetracks during the 2016 to 2019 seasons. The data were collected from the New York State Gaming Commission (NYSGC) Equine Breakdown, Death, Injury and Incident (EBDII), The Jockey Club InCompass Solutions, and The United States Trotting Association Pathway databases. Survival analysis was used to compare the predisposing factors by breed of the horse. The study included 431 horses (360 TB and 71 SB). There was a significant difference between the average incidence of attrition events per start for TB (0.304) and SB (0.035) horses. There was a significant difference in the median age to attrition between TB (4.2 years) and SB (7.5 years) horses. There was also a significant difference between the average number of starts to attrition for TB (13.6) and SB (125.5) horses. Among the causes of attrition events in the two breeds musculoskeletal injuries and medical conditions were significantly higher among TB horses, while sudden death was significantly higher among SB horses. The multivariate analysis showed that the hazard of attrition was affected by the age of the horse, age at first start, whether the horse was intact male, type of activity (racing training or other) and the breed of the horse. SB horses were less likely to experience attrition events in comparison to TB horses.

Serological evidence of West Nile viral infection in archived swine serum samples from Peninsular Malaysia.

West Nile virus (WNV), a neurotropic arbovirus, has been detected in mosquitos, birds, wildlife, horses, and humans in Malaysia, but limited information is available on WNV infection in Malaysian pigs. We tested 80 archived swine serum samples for the presence of WNV antibody and West Nile (WN) viral RNA using ID Screen West Nile Competition Multi-species enzyme-linked immunosorbent assay kits and WNV-specific primers in reverse transcription polymerase chain reaction assays, respectively. A WNV seroprevalence of 62.5% (50/80) at 95% confidence interval (51.6%-72.3%) was recorded, with a significantly higher seroprevalence among young pigs (weaner and grower) and pigs from south Malaysia. One sample was positive for Japanese encephalitis virus antibodies; WN viral RNA was not detected in any of the serum samples.

An Exploratory Study into the Implantation of Arytenoid Cartilage Scaffold in the Horse.

Respiratory function in the horse can be severely compromised by arytenoid chondritis, or arytenoid chondropathy, a pathologic condition leading to deformity and dysfunction of the affected cartilage. Current treatment in cases unresponsive to medical management is removal of the cartilage, which can improve the airway obstruction, but predisposes the patient to other complications like tracheal penetration of oropharyngeal content and dynamic collapse of the now unsupported soft tissue lateral to the cartilage. A tissue engineering approach to reconstructing the arytenoid cartilage would represent a significant advantage in the management of arytenoid chondritis. In this study, we explored if decellularized matrix could potentially be incorporated into the high motion environment of the arytenoid cartilages of horses. Equine arytenoid cartilages were decellularized and a portion of the resultant acellular scaffolds was implanted in a full-thickness defect created in the arytenoids of eight horses. The implantation was performed bilaterally in each horse, with one side randomly selected to receive an implant seeded with autologous bone marrow-derived nucleated cells (BMNCs). Arytenoids structure and function were monitored up to 4 months. In vivo assessments included laryngeal ultrasound, and laryngeal endoscopy at rest and during exercise on a high-speed treadmill. Histologic evaluation of the arytenoids was performed postmortem. Implantation of the cartilaginous graft had no adverse effect on laryngeal respiratory function or swallowing, despite induction of a transient granuloma on the medial aspect of the arytenoids. Ultrasonographic monitoring detected a postoperative increase in the thickness and cross-sectional area of the arytenoid body that receded faster in the arytenoids not seeded with BMNCs. The explanted tissue showed epithelialization of the mucosal surface, integration of the implant into the native arytenoid, with minimal adverse cellular reaction. Remodeling of the scaffold material was evident by 2 months after implantation. Preseeding the scaffold with BMNCs increased the rate of scaffold degradation and incorporation. Replacement of arytenoid portion with a tissue-engineered cartilaginous graft preseeded with BMNCs is surgically feasible in the horse, is well tolerated, and results in appropriate integration within the native tissue, also preventing laryngeal tissue collapse during exercise.

Evaluation of new leptospiral antigens for the diagnosis of equine leptospirosis: An approach using pan-genomic analysis, reverse vaccinology and antigenic selection.

BACKGROUND: The current gold standard diagnostic test for leptospirosis is the microscopic agglutination test (MAT), which has many drawbacks; therefore, the development of a better and easier serological test for leptospirosis is needed. OBJECTIVES: To apply reverse vaccinology (RV) and antigenic selection on the assortment of leptospiral targets and evaluate their potential for use as reagents for the diagnosis of equine leptospirosis. STUDY DESIGN: Cross-sectional study. METHODS: The antigenic selection parameters were: proteins with antigenicity score ≥0.5 (VaxiJen), at least one B cell epitope and size between 10 and 275 KDa. New leptospiral proteins were cloned, expressed and serologically screened against equine sera (n = 128) on a single analysis and comparative combinations. Sensitivity (Se) and specificity (Sp), accuracy, positive predictive value (PPV) and negative predictive value (NPV) were calculated. A BLAST with nucleotide and protein sequences was used to identify the serovar or species specificity. MAIN LIMITATIONS: This cross-sectional analysis had three main limitations: (a) The equine sera used in these tests were limited to sera submitted to the Animal Health Diagnosis Center and were only tested against seven serovars; (b) MAT results were considered being 'perfect', and the highest titre presented was considered being the infecting serovar, which may not hold true; (c) The strains used to represent the serovars and the limited number of different serovars and species included in the genetic analysis, which leads to the possibility that these proteins might be present in different species or serovars that perhaps would be seroprevalent in another geographic region. CONCLUSIONS: The new leptospiral antigens described in this research could increase the sensitivity and specificity of ELISA for detection of Leptospira exposure and the detection of leptospirosis in horses along with support from other clinical signs. Some of these new antigens might be used to improve the detection of infecting serovar.

Exposure to Zoonotic West Nile Virus in Long-Tailed Macaques and Bats in Peninsular Malaysia.

The role of wildlife such as wild birds, macaques, and bats in the spreading and maintenance of deadly zoonotic pathogens in nature have been well documented in many parts of the world. One such pathogen is the mosquitoes borne virus, namely the West Nile Virus (WNV). Previous research has shown that 1:7 and 1:6 Malaysian wild birds are WNV antibody and RNA positive, respectively, and bats in North America may not be susceptible to the WNV infection. This study was conducted to determine the status of WNV in Malaysian macaques and bats found in mangrove forests and caves, respectively. Archive sera and oropharyngeal swabs from long-tailed macaques were subjected to the antibody detection using WNV competitive enzyme-linked immunosorbent assay (c-ELISA) and WNV RNA using RT-PCR, respectively, while the archive oropharyngeal and rectal swabs from bats were subjected to RT-PCR without serological analysis due to the unavailability of serum samples. The analysis revealed a WNV seropositivity of 29.63% (24/81) and none of the macaques were positive for WNV RNA. Meanwhile, 12.2% (5/41) of the bats from Pteropodidae, Emballonuridae, and Rhinolophidae families tested positive for WNV RNA. Here, we show a high WNV antibody prevalence in macaques and a moderate WNV RNA in various Malaysian bat species, suggesting that WNV circulates through Malaysian wild animals and Malaysian bat species may be susceptible to the WNV infection.

Evidence of West Nile virus infection in migratory and resident wild birds in west coast of peninsular Malaysia.

West Nile virus (WNV) is a zoonotic mosquito-borne flavivirus that is harbored and amplified by wild birds via the enzootic transmission cycle. Wide range of hosts are found to be susceptible to WNV infection including mammals, amphibians and reptiles across the world. Several studies have demonstrated that WNV was present in the Malaysian Orang Asli and captive birds. However, no data are available on the WNV prevalence in wild birds found in Malaysia. Therefore this study was conducted to determine the serological and molecular prevalence of WNV in wild birds in selected areas in the West Coast of Peninsular Malaysia. Two types of wild birds were screened, namely migratory and resident birds in order to explore any possibility of WNV transmission from the migratory birds to the resident birds. Thus, a cross-sectional study was conducted at the migratory birds sanctuary located in Kuala Gula, Perak and Kapar, Selangor by catching 163 migratory birds, and 97 resident birds from Kuala Gula and Parit Buntar, Perak at different time between 2016 and 2017 (Total, n = 260). Blood and oropharyngeal swabs were collected for serological and molecular analysis, respectively. Serum were screened for WNV antibodies using a commercial competitive ELISA (c-ELISA) (ID Screen® West Nile Competition Multi-species ELISA, ID VET, Montpellier, France) and cross-reactivity towards Japanese Encephalitis virus (JEV) was also carried out using the JEV-double antigen sandwich (DAS) ELISA. Oropharyngeal swabs were subjected to one-step RT-PCR to detect WNV RNA, in which positive reactions were subsequently sequenced. WNV seropositive rate of 18.71% (29/155) at 95% CI (0.131 to 0.260) and molecular prevalence of 15.2% (16/105) at 95% CI (0.092 to 0.239) were demonstrated in migratory and resident wild birds found in West Coast Malaysia. Phylogenetic analyses of the 16 WNV isolates found in this study revealed that the local strains have 99% similarity to the strains from South Africa and were clustered under lineage 2. Evidence of WNV infection in resident and migratory birds were demonstrated in this study. As a summary, intervention between migratory birds, resident birds and mosquitoes might cause the introduction and maintenance of WNV in Malaysia, however the assumption could be further proven by studying the infection dynamics in the mosquitoes present in the studied areas.

Management of postoperative dysphagia after prosthetic laryngoplasty or arytenoidectomy.

OBJECTIVE: To evaluate the efficacy of various treatments for dysphagia after laryngeal surgery. STUDY DESIGN: Retrospective. ANIMALS: Horses treated for dysphagia after laryngeal surgery. METHODS: Medical records of horses treated for dysphagia after prosthetic laryngoplasty (PLP) or partial arytenoidectomy (PA) were reviewed. Signalment, prior surgery, preoperative videoendoscopic diagnosis, and surgical treatments were recorded. Short- and long-term follow-up were obtained. Chi-square and logistical regression were performed to correlate independent variables and outcomes. RESULTS: Forty-four percent of horses with prior PLP and 88% of horses with prior PA had persistent resting dorsal displacement of the soft palate (DDSP). Vocal fold augmentation (VFA) as a single treatment was most commonly performed (n = 22), followed by laryngoplasty removal (LPR; n = 1). Fifteen horses received a combination of treatments: VFA (n = 15), LPR (n = 4), laryngeal tie-forward (LTF; n = 2), and esophageal release (n = 1). Short-term clinical outcomes were reported; 80% of horses treated with VFA (n = 35) had resolution of dysphagia, and 20% were improved. Among the 33 horses available to long-term follow-up, 31 (94%) returned to some level of work, and DDSP during work was reportedly resolved in 23 of 25 horses. CONCLUSION: Vocal fold augmentation resulted in resolution of symptoms relating to dysphagia in 81% of horses. Twenty percent of horses treated with LPR had sustained resolution of dysphagia. A combination of LPR, VFA, LTF was performed in 18% of horses. CLINICAL SIGNIFICANCE: Treatment options directed at altering laryngeal geometry and/or position of the larynx were successful in resolving 86% of dysphagia in horses previously regarded as very difficult to treat.