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  1. Khor KH, Chin MX
    J Adv Vet Anim Res, 2020 Sep;7(3):501-508.
    PMID: 33005676 DOI: 10.5455/javar.2020.g446
    Objective: Annual health screening inclusive of heart workup is recommended for the detection of heart diseases, especially in asymptomatic patients (no clinical signs). This study determined the occurrences of the common heart disease and the risk factors in apparently healthy cats.

    Materials and methods: This prospective study that screened 59 healthy cats and the status of the heart were evaluated based on a combination of findings from physical examination, electrocardiography, blood pressure measurement, routine blood test, urinalysis, and total thyroid level.

    Results: Approximately 40.7% (n = 24/59) of the apparently healthy cats were diagnosed with heart disease hypertrophic cardiomyopathy (62.5%) remains to be the most commonly diagnosed. The mean age was 4.9-year old (age range, 7-month-old to 19-year-old). The prevalence was higher in males (45.0%; n = 17/38) cats, especially the domestic shorthairs (46.0%; n = 11/24). Among the healthy cats with vertebral heart scale (VHS) > 8.0, only 52% (n = 12/23) of them were diagnosed with cardiomyopathy. However, 33% (n = 12/36) of the cats with normal VHS ≤ 7.9 were diagnosed with heart disease. Consistently, all healthy cats with abnormal heart sounds were diagnosed with heart disease. About 31.4% (n = 16/51) of these cats with typical heart sound had cardiomyopathy too.

    Conclusion: The occurrence of cardiomyopathy in apparently healthy cats has no association with the patient's age, sex, and VHS, except for the heart sound. Echocardiography remains the best diagnostic tool, as normal heart size and normal heart sound do not exclude cardiomyopathy in this group of apparently healthy cats.

  2. Khor KH, Campbell FE, Owen H, Shiels IA, Mills PC
    Vet J, 2015 Feb;203(2):161-8.
    PMID: 25573453 DOI: 10.1016/j.tvjl.2014.11.018
    The histological features of feline hypertrophic cardiomyopathy (HCM) have been well documented, but there are no reports describing the histological features in mild pre-clinical disease, since cats are rarely screened for the disease in the early stages before clinical signs are apparent. Histological changes at the early stage of the disease in pre-clinical cats could contribute to an improved understanding of disease aetiology or progression. The aim of this study was to evaluate the histological features of HCM in the left ventricular (LV) myocardium of cats diagnosed with pre-clinical HCM. Clinically healthy cats with normal (n = 11) and pre-clinical HCM (n = 6) were identified on the basis of echocardiography; LV free wall dimensions (LVFWd) and/or interventricular septal wall (IVSd) dimensions during diastole of 6-7 mm were defined as HCM, while equivalent dimensions <5.5 mm were defined as normal. LV myocardial sections were assessed and collagen content and inflammatory cell infiltrates were quantified objectively. Multifocal areas of inflammatory cell infiltration, predominantly lymphocytes, were observed frequently in the left myocardium of cats with pre-clinical HCM. Tissue from cats with pre-clinical HCM also had a higher number of neutrophils and a greater collagen content than the myocardium of normal cats. The myocardium variably demonstrated other features characteristic of HCM, including arteriolar mural hypertrophy and interstitial fibrosis and, to a lesser extent, myocardial fibre disarray and cardiomyocyte hypertrophy. These results suggest that an inflammatory process could contribute to increased collagen content and the myocardial fibrosis known to be associated with HCM.
  3. Khor KH, Moore TA, Shiels IA, Greer RM, Arumugam TV, Mills PC
    PLoS One, 2016;11(1):e0146022.
    PMID: 26727203 DOI: 10.1371/journal.pone.0146022
    PURPOSE: Inflammation may contribute to the pathogenesis of specific cardiovascular diseases, but it is uncertain if mediators released during the inflammatory process will affect the continued efficacy of drugs used to treat clinical signs of the cardiac disease. We investigated the role of the complement 5a receptor 1 (C5aR1/CD88) in the cardiac response to inflammation or atenolol, and the effect of C5aR1 deletion in control of baseline heart rate in an anesthetized mouse model.

    METHODS: An initial study showed that PMX53, an antagonist of C5aR1 in normal C57BL6/J (wild type, WT) mice reduced heart rate (HR) and appeared to have a protective effect on the heart following induced sepsis. C5aR1 knockout (CD88-/-) mice had a lower HR than wild type mice, even during sham surgery. A model to assess heart rate variability (HRV) in anesthetized mice was developed to assess the effects of inhibiting the β1-adrenoreceptor (β1-AR) in a randomized crossover study design.

    RESULTS: HR and LF Norm were constitutively lower and SDNN and HF Norm constitutively higher in the CD88-/- compared with WT mice (P< 0.001 for all outcomes). Administration of atenolol (2.5 mg/kg) reduced the HR and increased HRV (P< 0.05, respectively) in the wild type but not in the CD88-/- mice. There was no shift of the sympathovagal balance post-atenolol in either strains of mice (P> 0.05), except for the reduced LF/HF (Lower frequency/High frequency) ratio (P< 0.05) at 60 min post-atenolol, suggesting increased parasympathetic tone of the heart due to the effect of atenolol administration. The HR of the WT mice were lower post atenolol compared to the CD88-/- mice (P = 0.001) but the HRV of CD88-/- mice were significantly increased (P< 0.05), compared with WT mice.

    CONCLUSION: Knockout of the C5aR1 attenuated the effect of β1-AR in the heart, suggesting an association between the β1-AR and C5aR1, although further investigation is required to determine if this is a direct or causal association.

  4. Rahman SA, Khor KH, Khairani-Bejo S, Lau SF, Mazlan M, Roslan A, et al.
    J Vet Diagn Invest, 2021 Sep;33(5):834-843.
    PMID: 34148436 DOI: 10.1177/10406387211024575
    Leptospirosis is a serious bacterial disease that affects both humans and animals. A wide range of symptoms have been described in humans; the disease in dogs is commonly associated with kidney and/or liver disease. In Malaysia, information about the common serovars infecting dogs is limited. Therefore, we investigated the occurrences of leptospirosis in 124 pet dogs diagnosed with kidney and/or liver disease. Blood, urine, abdominal effusion, and/or kidney and liver were collected from the dogs. Based on microscopic agglutination testing, 53 of 124 (42.7%) dogs were seropositive for leptospiral exposure. Sera were frequently positive to serovars Bataviae (n = 12), Javanica (n = 10), and Icterohaemorrhagiae (n = 10). Direct detection using PCR showed that 42 of 124 (33.9%) of the whole blood and 36 of 113 (31.9%) urine samples were positive for pathogenic Leptospira spp. By PCR, 2 of 23 (9.1%) kidney and 2 of 23 (9.1%) liver were positive for pathogenic Leptospira spp. Abdominal effusion from 4 dogs were PCR-positive for pathogenic Leptospira spp. The species detected were L. interrogans, L. borgpetersenii, L. kirschneri, and L. kmetyi by partial 16S rRNA sequencing. We further identified and characterized 11 Leptospira spp. isolates from 8 dogs as serovars Bataviae, Javanica, and Australis. The mortality rate of the Leptospira-infected dogs was high (18 of 53; 34%).
  5. Khor KH, Shiels IA, Campbell FE, Greer RM, Rose A, Mills PC
    Vet J, 2014 Feb;199(2):229-35.
    PMID: 24321367 DOI: 10.1016/j.tvjl.2013.11.006
    Analysis of heart rate (HR) and heart rate variability (HRV) are powerful tools to investigate cardiac diseases, but current methods, including 24-h Holter monitoring, can be cumbersome and may be compromised by movement artefact. A commercially available data capture and analysis system was used in anaesthetised healthy cats to measure HR and HRV during pharmacological manipulation of HR. Seven healthy cats were subjected to a randomised crossover study design with a 7 day washout period between two treatment groups, placebo and atenolol (1mg/kg, IV), with the efficacy of atenolol to inhibit β1 adrenoreceptors challenged by epinephrine. Statistical significance for the epinephrine challenge was set at P<0.0027 (Holm-Bonferroni correction), whereas a level of significance of P<0.05 was set for other variables. Analysis of the continuous electrocardiography (ECG) recordings showed that epinephrine challenge increased HR in the placebo group (P=0.0003) but not in the atenolol group. The change in HR was greater in the placebo group than in the atenolol group (P=0.0004). Therefore, compared to cats pre-treated with placebo, pre-treatment with atenolol significantly antagonised the tachycardia while not significantly affecting HRV. The increased HR in the placebo group following epinephrine challenge was consistent with a shift of the sympathovagal balance towards a predominantly sympathetic tone. However, the small (but not significant at the critical value) decrease in the normalised high-frequency component (HFnorm) in both groups of cats suggested that epinephrine induced a parasympathetic withdrawal in addition to sympathetic enhancement (increased normalised low frequency component or LFnorm). In conclusion, this model is a highly sensitive and repeatable model to investigate HRV in anaesthetised cats that would be useful in the laboratory setting for short-term investigation of cardiovascular disease and subtle responses to pharmacological agents in this species.
  6. Abdul Rahman MS, Khor KH, Khairani-Bejo S, Lau SF, Mazlan M, Roslan MA
    Animals (Basel), 2021 Nov 29;11(12).
    PMID: 34944181 DOI: 10.3390/ani11123405
    Canine leptospirosis is commonly associated with kidney and/or liver disease. It has been widely reported and causes public health concerns due to its zoonotic potential and its re-emergence, resulting from close contact between humans and dogs. The current study identified potential risk and predictive factors for dogs diagnosed with kidney and/or liver disease due to leptospirosis. A total of 124 client-owned dogs were recruited, and information such as signalment, medical history, management, and clinical findings were documented. Samples collected from the recruited dogs were directly tested using polymerase chain reaction (PCR) and subsequently inoculated for bacterial isolation. Statistical analyses were descriptively analyzed, and risk analyses were performed using Pearson chi-square tests and logistic regression. A total of 53 dogs (42.7%) were positive for leptospiral infection based on PCR, and 10 leptospiral isolates were successfully recovered from eight dogs. The mortality rate of infected dogs was 34.0% (18/53). Medium and large dog breeds, with a history of exposure to rats, and managed outdoors had a greater risk for leptospirosis (p < 0.05). The significant predictors for the dogs' positivity were the presence of rats and acute clinical illness (p < 0.05). Administration of antibiotics and the detection of clinical illness at an early stage of the disease improved the survivability of the dogs (p < 0.05). Identifying the profile of dogs that are at risk to leptospirosis could be useful in the design of diagnostic and treatment strategies, as well as to increase awareness for prevention of the disease.
  7. Mohd Isa NH, Selvarajah GT, Khor KH, Tan SW, Manoraj H, Omar NH, et al.
    Vet Microbiol, 2019 Sep;236:108382.
    PMID: 31500720 DOI: 10.1016/j.vetmic.2019.08.005
    Feline morbillivirus (FeMV), a novel virus from the family of Paramyxoviridae, was first identified in stray cat populations. The objectives of the current study were to (i) determine the molecular prevalence of FeMV in Malaysia; (ii) identify risk factors associated with FeMV infection; and (iii) characterise any FeMV isolates by phylogenetic analyses. Molecular analysis utilising nested RT-PCR assay targeting the L gene of FeMV performed on either urine, blood and/or kidney samples collected from 208 cats in this study revealed 82 (39.4%) positive cats. FeMV-positive samples were obtained from 63/124 (50.8%) urine and 20/25 (80.0%) kidneys while all blood samples were negative for FeMV. In addition, from the 35 cats that had more than one type of samples collected (blood and urine; blood and kidney; blood, urine and kidney), only one cat had FeMV RNA in the urine and kidney samples. Risk factors such as gender, presence of kidney-associated symptoms and cat source were also investigated. Male cats had a higher risk (p = 0.031) of FeMV infection than females. In addition, no significant association (p = 0.083) was observed between the presence of kidney-associated symptoms with FeMV status. From the 82 positive samples, FeMV RNA was detected from 48/82 (58.5%) pet cats and 34/126 (27.0%) shelter cats (p 
  8. Alashraf AR, Lau SF, Khor KH, Khairani-Bejo S, Bahaman AR, Roslan MA, et al.
    Top Companion Anim Med, 2019 Mar;34:10-13.
    PMID: 30808490 DOI: 10.1053/j.tcam.2018.12.002
    Leptospirosis is one of the most widespread zoonotic diseases and despite extensive research, there is still a paucity of information regarding this disease in cats. The aim of this study was to investigate the prevalence of leptospirosis among the shelter cat population in Malaysia and to determine the most common infective Leptospira serogroups among them. Blood samples were collected from a total of 110 cats from 4 different shelters. The sampled cats appeared healthy, with minimal evidence of feline upper respiratory disease. The Microscopic Agglutination Test was used to detect anti-Leptospira antibodies against 20 pathogenic serovars. Based on a cut-off antibody titer of ≥1:100, 20 of 110 sheltered cats, showed presence of anti-Leptospira antibodies against at least 1 serovar. The serodetection of leptospirosis was 18.18% (95% confidence interval 12.09-26.42). The most commonly detected serogroups were Bataviae, Javanica, and Ballum, with antibody titers ranging from 1:100 to 1:1600. Knowledge of the predominant infective serovars in hosts worldwide and regionally is imperative for understanding the epidemiology of this zoonotic disease. Serosurveillance is the first step in this process. Further studies are warranted for investigation of urinary shedding in naturally infected cats with leptospirosis, using Polymerase chain reaction (PCR) and organism isolation followed by serovars identification.
  9. Goh SH, Khor KH, Radzi R, Lau SF, Khairani-Bejo S, Abdul Rahman MS, et al.
    Top Companion Anim Med, 2021 Nov;45:100562.
    PMID: 34271178 DOI: 10.1016/j.tcam.2021.100562
    Leptospirosis is an endemic zoonoses of global proportions. Stray dogs have been postulated to play a role in disease transmission; however, supporting information are still limited. Roaming behavior may not only predispose the dogs to infection, but could also contribute to disease spread. In this study, the susceptibility of urban stray dogs in shedding Leptospira spp. was determined. Blood, urine, and tissue samples of kidney and liver were collected from 100 dogs from 2 animal control facilities. Serological testing using microscopic agglutination test (MAT) were performed on blood against 20 leptospiral serovars with a cut-off titre of ≥ 1:100. Samples were cultured onto semi-solid Ellinghausen and McCullough modified by Johnson and Harris (EMJH) media. Isolates were identified using molecular polymerase chain reaction (PCR) using 2 primers (16s rRNA and LipL32) and hyperimmune serum (HIS) MAT. The seroprevalence for the dogs positive for leptospirosis was 32% (n=32/100) with the following detected serovars: Javanica (n=13), Bataviae (n=10), Icterohaemorrhagiae (n=3), Autumnalis (n=2), Canicola (n=1), Pyrogenes (n=1), Copenhageni (n=1), and Australis (n=1). Six Leptospira spp. isolated were procured from urine (n=2), kidney (n=2) and liver (n=2). All 6 isolates belonged to L. interrogans, a pathogenic variant of Leptospira spp. Serotyping and phylogenetic analysis suggested serovar Bataviae (n=5) and serovar Canicola (n=1). Presence of vaccinal serovars (Icterohaemorrhagiae and Canicola) suggested potential post-vaccination antibodies but the predominance of non-vaccinal serovars (Javanica and Bataviae) indicate the possibility of current infection or post-exposure. Isolation of Leptospira spp. directly from urine sample not only suggested an active infection but highlighted the potential shedding capability among these stray dogs. These findings further strengthen speculations that urban stray dogs could play a role in transmission and dissemination of leptospirosis through their constant movement. The urine of infected dogs may contaminate the environment, posing a major public health threat.
  10. Lau SF, Wong JY, Khor KH, Roslan MA, Abdul Rahman MS, Bejo SK, et al.
    Top Companion Anim Med, 2017 Dec;32(4):121-125.
    PMID: 29525230 DOI: 10.1053/j.tcam.2017.12.001
    Working dogs are canine animals that have been trained to assist human beings in carrying out various tasks. They help in guarding property, performing rescues, assisting the visually impaired or physically handicapped, searching for drugs, explosives, and others. Leptospirosis is one of the most widespread zoonotic diseases in the world and a commonly occurring disease of the tropics and subtropics. In Malaysia, all working dogs are normally vaccinated with serovars, Pomona, Icterohaemorrhagiae, Canicola, and Grippotyphosa based on protocols recommended from other countries. The duration of immunity in vaccinated dogs for Leptospira can last up to 13 months; however, there is no full crossprotection between the different serovars. Five representative canine units from different government agencies in Malaysia (n = 96 dogs) were recruited in this study. For detection, the microscopic agglutination test was performed by incubating the serum from dogs with various serovars of leptospires, namely, Icterohaemorrhagiae, Canicola, Pomona, Grippotyphosa, Australis, Bataviae, Javanica, Tarassovi, Hebdomadis, Lai, and Pyrogenes. The plasma obtained was used for polymerase chain reaction (PCR) analysis, for the detection of 16S rRNA, and lipL 32 genes of Leptospira. Out of the 96 dogs sampled, only 3 dogs were positive toward serovars, Australis, Bataviae, and Javanica, based on the cutoff point at 1:80. The seroprevalence of canine leptospirosis in this population was 3.1% (n = 3/96). However, all 96 blood samples of working dogs tested negative for both pathogenic and nonpathogenic Leptospira genes. The results revealed that, by vaccination alone, working dogs were not fully protected against leptospirosis and could pose a risk to dog handlers. A preventative and control protocol for leptospirosis is warranted, and its implementation should be monitored and improved accordingly from time to time, in order to maintain a healthy condition in both working dogs and their handlers.
  11. Rahman MSA, Khor KH, Khairani-Bejo S, Lau SF, Mazlan M, Roslan MA, et al.
    J Vet Res, 2023 Jun;67(2):187-195.
    PMID: 38143826 DOI: 10.2478/jvetres-2023-0024
    INTRODUCTION: Canine leptospirosis has always been a differential diagnosis in dogs presenting with clinical signs and blood profiles associated with kidney and/or liver disease. The conventional polymerase chain reaction (PCR) provides diagnoses, but real-time PCR-based tests provide earlier confirmation and determine the severity of infection, especially in the acute stage, allowing early detection for immediate treatment decisions. To our knowledge, real-time PCR has not been routinely adopted for clinical investigation in Malaysia. This study evaluated TaqMan real-time PCR (qPCR) assays diagnosing leptospirosis and compared their applicability to clinical samples from dogs with kidney and/or liver disease against a conventional PCR reference.

    MATERIAL AND METHODS: The qPCR assays were validated using existing leptospiral isolates. Whole blood and urine samples were analysed using a conventional PCR, LipL32(1) and LipL32(2) qPCRs and a microscopic agglutination test. The sensitivity and specificity of the qPCRs were determined.

    RESULTS: The LipL32(1) qPCR assay had more diagnostic value than the LipL32(2) qPCR assay. Further evaluation of this assay revealed that it could detect as low as five DNA copies per reaction with high specificity for the tested leptospiral strains. No cross-amplification was observed with other organisms. Analysing the clinical samples, the LipL32(1) qPCR assay had 100.0% sensitivity and >75.0% specificity.

    CONCLUSION: The LipL32(1) qPCR assay is sensitive, specific and has the potential to be applied in future studies.

  12. Alashraf AR, Lau SF, Khairani-Bejo S, Khor KH, Ajat M, Radzi R, et al.
    PLoS One, 2020;15(3):e0230048.
    PMID: 32155209 DOI: 10.1371/journal.pone.0230048
    Leptospirosis is one of the most widespread zoonotic diseases and can infect both humans and animals worldwide. Healthy cat, as a potential source of exposure to humans, are likely underestimated owing to the lack of overt clinical signs associated with Leptospira spp. infection in this species. The aim of the study was to determine the exposure, shedding, and carrier status of leptospires in shelter cats in Malaysia by using serological, molecular, and bacteriological methods. For this study, 82 healthy cats from two shelters were sampled. The blood, urine, and kidneys were tested using the microscopic agglutination test (MAT), polymerase chain reaction (PCR), and bacterial culture. On the basis of serological, molecular, and/or culture techniques, the total detection of leptospiral infection was 29.3% (n = 24/82). Through culture techniques, 16.7% (n = 4/24) of the cats that tested positive were carriers with positive kidney cultures, and one cat was culture positive for both urine and kidney. The Leptospira spp. isolates were identified as pathogenic L. interrogans serovar Bataviae through serological and molecular methods. Through serological techniques, 87.5% (n = 21/24) had positive antibody titers (100-1600) and most of the Bataviae serogroup (n = 19/21). Using PCR, 16.7% (n = 4/24) of cats were shown to have pathogenic Leptospira spp. DNA in their urine. Furthermore, three out of four culture positive cats were serology negative. The present study reports the first retrieval of pathogenic leptospires from urine and kidneys obtained from naturally infected cats. The results provide evidence of the potential role of naturally infected cats in the transmission of leptospires. Additionally, leptospiral infection occurs sub-clinically in cats. The culture isolation provides evidence that healthy cats could be reservoirs of leptospiral infection, and this information may promote the development of disease prevention strategies for the cat population.
  13. Goh SH, Khor KH, Ismail R, Megat Abdul Rani PA, Mohd Mohidin TB, Bahaman AR, et al.
    Trop Biomed, 2020 Dec 01;37(4):1074-1082.
    PMID: 33612759 DOI: 10.47665/tb.37.4.1074
    The incidence of leptospirosis seems to be on the rise and could be an alarming indirect indication of a global re-emergence. It is a potential public health threat when dogs are speculated to be involved in the transmission of leptospirosis through possible subclinical harbouring of Leptospira spp. and subsequent shedding into the environment. This study aimed to detect anti-leptospiral antibodies among dogs and their handlers using the microscopic agglutination test (MAT). Blood samples from 266 apparently healthy dogs and 194 dog handlers were collected at four working dog organisations and four dog shelters. Serum samples were tested using MAT against 20 leptospiral serovars with a cut-off titre >=1:100 (dog) and >=1:50 (dog handlers). Seventy dogs (70/266; 26.3%) were seropositive mainly against serovars Icterohaemorrhagiae, Ballum, Bataviae and Javanica (titres ranged: 1:100-1:800). Sixty-seven dog handlers (67/194; 34.5%) were seropositive mainly against serovars Grippotyphosa, Icterohaemorrhagiae and Malaysia (titres ranged: 1:50-1:200). Dogs were seropositive due to exposure, vaccination or active infection. Seropositive dog handlers could indicate exposure or active infection. This shows the potential of dogs in maintaining and spreading the infection in Malaysia. Due to the occupational risk as a result of frequent contact with dogs and exposure to contaminated environments, dog handlers should be made aware of the presence of this zoonotic disease.
  14. Goh SH, Ismail R, Lau SF, Megat Abdul Rani PA, Mohd Mohidin TB, Daud F, et al.
    PMID: 31035316 DOI: 10.3390/ijerph16091499
    This study determined the potential risk factors that may contribute to seropositivity among dogs and dog handlers from working dog and dog shelter institutions. Data was collected from dogs (n = 266) and dog handlers (n = 161) using a standardised guided questionnaire. Serum obtained from the dogs and dog handlers was tested using the microscopic agglutination test (MAT). A logistic regression analysis was used to predict leptospiral seropositivity of dogs and dog handlers based on potential risk factors. A total of 22.2% of dogs and 21.7% of dog handlers were seropositive. The significant predictors for the dogs' seropositivity were presence of rats (OR = 4.61 (95% CI: 1.05, 20.33), p = 0.043) and shared common area (OR = 5.12 (95% CI: 1.94, 13.46), p = 0.001) within the organisation. Significant predictor for dog handler seropositivity was contact time with the dogs of more than six hours/day (OR = 3.28 (95% CI: 1.28, 8.40), p = 0.013) after controlling for the effect of other risk factors such as small mammal contact, rat infestation at home, flooding at housing area (within three months) and urban locality. The exposure to various disease sources identified poses risk to dogs and dog handlers. Risk could be reduced with adequate application of protection at work while handling dogs and thus limiting contact with these sources and reducing exposure to infection.
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