Rapid detection of equine infectious anaemia virus nucleic acid by insulated isothermal RT-PCR assay to aid diagnosis under field conditions.

BACKGROUND: Control of equine infectious anaemia (EIA) currently depends on serological diagnosis of infected equids. However, recently infected equids may not produce detectable anti-EIAV antibodies up to 157 days post infection and so present a high transmission risk. Therefore, direct nucleic acid detection methods are urgently needed to improve EIAV surveillance and management programs in counties where the disease is endemic. OBJECTIVES: To evaluate a field-deployable, reverse transcription-insulated isothermal PCR (RT-iiPCR) assay targeting the conserved 5' untranslated region (5' UTR)/exon 1 of the tat gene of EIAV. STUDY DESIGN: The analytical and clinical performance of the newly developed EIAV RT-iiPCR was evaluated by comparison with a EIAV real-time RT-PCR (RT-qPCR) along with the AGID test. METHODS: Analytical sensitivity was determined using in vitro transcribed RNA containing the target area of the 5' UTR/tat gene and samples from two EIAV-positive horses. Specificity was verified using nine common equine viruses. Clinical performance was evaluated by comparison with EIAV RT-qPCR and AGID using samples derived from 196 inapparent EIAV carrier horses. RESULTS: EIAV RT-iiPCR did not react with other commonly encountered equine viruses and had equivalent sensitivity (95% detection limit of eight genome equivalents), with a concordance of 95.41% to conventional EIAV RT-qPCR. However, the RT-qPCR and RT-iiPCR had sensitivities of 43.75 and 50.00%, respectively, when compared to the AGID test. MAIN LIMITATIONS: Low viral loads commonly encountered in inapparent EIAV carriers may limit the diagnostic sensitivity of RT-PCR-based tests. CONCLUSIONS: Although EIAV RT-iiPCR is not sufficiently sensitive to replace the current AGID test, it can augment control efforts by identifying recently exposed or "serologically silent" equids, particularly as the latter often represent a significant transmission risk because of high viral loads. Furthermore, the relatively low cost and field-deployable design enable utilisation of EIAV RT-iiPCR even in remote regions.

On-site detection of equid alphaherpesvirus 3 in perineal and genital swabs of mares and stallions.

Equine coital exanthema (ECE) is an infectious, venereally transmitted muco-cutaneous disease affecting mares and stallions, caused by equid alphaherpesvirus 3 (EHV3). Diagnostic tools for rapid identification of EHV3 are of primary importance to diminish the risk of EHV3 dissemination at the time of breeding. In the last years, it has been shown that the performance of the insulated-isothermal polymerase chain reaction (iiPCR) is comparable to virus isolation, nested PCR and real-time PCR (qPCR) in detecting pathogens of various animal species. Analytical sensitivity and specificity of the iiPCR were compared with a qPCR, using a plasmid containing the target region of the EHV3 glycoprotein G gene and an Argentinian EHV3 isolate (E/9283/07 C3A). In order to evaluate the diagnostic performance of the iiPCR, nucleic acids of 85 perineal and genital swabs (PGS) of mares and stallions were extracted by tacoTM mini and tested by both techniques. EHV3 was detected in 46 and 45 of the 85 PGS by the iiPCR and qPCR, respectively. There was almost perfect agreement between the two diagnostic methods (98.82%; 95% CI: 95.03-100%; κ = 0.98). The iiPCR had a limit of detection of 95.00% at 6 genome equivalents per reaction and a detection endpoint for viral DNA comparable to that of the qPCR, and did not react with six non-targeted equine pathogens. The iiPCR represents a sensitive and specific method for the rapid on-site diagnosis of EHV3 infection. Its routinely implementation in breeding facilities, and artificial insemination and embryo transfer centers, will contribute to prevent the dissemination of this venereal, highly contagious disease in horses.

Validation of a KHV antibody enzyme-linked immunosorbent assay (ELISA).

Koi herpesvirus (KHV) causes KHV disease (KHVD). The virus is highly contagious in carp or koi and can induce a high mortality. Latency and, in some cases, a lack of signs presents a challenge for virus detection. Appropriate immunological detection methods for anti-KHV antibodies have not yet been fully validated for KHV. Therefore, it was developed and validated an enzyme-linked immunosorbent assay (ELISA) to detect KHV antibodies. The assay was optimized with respect to plates, buffers, antigens and assay conditions. It demonstrated high diagnostic and analytical sensitivity and specificity and was particularly useful at the pond or farm levels. Considering the scale of the carp and koi industry worldwide, this assay represents an important practical tool for the indirect detection of KHV, also in the absence of clinical signs.

A Rapid Field-Deployable Reverse Transcription-Insulated Isothermal Polymerase Chain Reaction Assay for Sensitive and Specific Detection of Bluetongue Virus.

Bluetongue is a non-contagious, haemorrhagic, Culicoides-borne disease of ruminants. The causative agent, bluetongue virus (BTV), is a member of the Orbivirus genus of the Reoviridae family. So far, 26 BTV serotypes have been identified worldwide. The global distribution of bluetongue has been expanding, and rapid detection of BTV, preferably in the field, is critical for timely implementation of animal movement restrictions and vector control measures. To date, many laboratory-based, molecular assays for detection of BTV have been developed. These methods require the samples to be shipped to a central laboratory with sophisticated instruments and highly skilled technicians to perform the assays, conduct analyses and interpret the results. Here, we report the development and evaluation of a rapid, portable, user-friendly, pan-BTV reverse transcription-insulated isothermal polymerase chain reaction (RT-iiPCR) assay that can potentially be used in low-resource field conditions. The total length of the assay was <60 min, and at the end of the assay, the results were automatically displayed as '+' or '-' without the need for data interpretation. The RT-iiPCR assay detected 36 BTV isolates and two in vitro transcribed RNA samples representing all 26 BTV serotypes. The assay did not cross-react with other animal viruses tested, including two closely related orbiviruses. The analytical sensitivity of the assay was as low as nine copies of in vitro transcribed double-stranded BTV RNA. Analysis of BTV-infected whole blood samples showed that the BTV RT-iiPCR assay was as sensitive as real-time RT-PCR. The assay can potentially be used for rapid screening of animals for BTV in routine diagnostics and for monitoring bluetongue outbreaks both in ruminants and in Culicoides vectors in the field and in the laboratory.

Effects of dietary recombinant chlorella supplementation on growth performance, meat quality, blood characteristics, excreta microflora, and nutrient digestibility in broilers.

The use of chlorella as an immune stimulant to enhance nonspecific host defense mechanisms or as an antimicrobial to inhibit bacterial growth has been reported. Thus, the aim of the present study was to clarify the effect of recombinant chlorella supplementation on growth performance, meat quality, and the blood profile, excreta microflora, and nutrient digestibility in broilers. A total of 375 one-day-old ROSS 308 broilers (male and female) were allotted to 5 dietary treatments using 5 cages with 15 chicks per cage. Treatments were: 1) NC, basal diet supplemented with 1.0% E. coli fermented liquor (EFL); 2) PC1, 0.2% EFL with chlorella; 3) PC2, 1.0% EFL with chlorella; 4) T1, 0.2% EFL with chlorella (anti-viral); and 5) T2, 1.0% EFL with chlorella (anti-viral). The broilers in the T2 treatment groups showed higher body weight gain (BGW) by 2.55% (P < 0.01) and lower feed conversion ratio (FCR) by 2.75% (P < 0.05) compared with those fed the control NC treatment group. Moreover, the blood contents of blood urea nitrogen (BUN), creatinine, and IgA in the broilers of the T2 treatment group were significantly increased by 28.12, 23.07, and 29.72%, respectively -more than those found in the broilers of the NC treatment group (P < 0.01). In contrast, the LDL/C in the blood from the animals in the T2 treatment group was significantly decreased by 23.23% - more than that in the blood from the NC broilers (P < 0.05). Based on these results, we suggest that the dietary supplementation of broilers with recombinant chlorella could improve their growth performance, increase the concentration of IgA and apparently metabolizable nitrogen in the blood, and decrease ammonia emissions. Therefore, our findings have important implications for the effect of recombinant chlorella supplementation through increasing the concentration of IgA and the level of metabolizable nitrogen.

Post-treatment impact and needs of prostate cancer survivors in Malaysia; a qualitative study

Introduction and Objectives There are limited studies conducted on the needs of cancer survivorsin developing countries like Malaysia. This qualitative study aimed at exploring the post-treatmentimpact and needs of prostate cancer survivors.Method: A qualitative study design was used. One in-depth interview and four focus groupdiscussions were conducted with 24 prostate cancer survivors (age range: 58–79 years) fromgovernment and private hospitals in Malaysia in 2013. Trained researchers used a topic guide toguide the interviews, which were audio-recorded, transcribed verbatim, checked and managed withNvivo 10 software. A thematic approach was used to analyse the data.Result: Three main themes emerged from the analysis: (a) impact of prostate cancer on thesurvivors, (b) support needed for coping and (c) information needs. Prostate cancer has animportant impact on the survivors’ lifestyle after treatment. Some of them have to live with thepost-treatment side effects. They were anxious about the possibility of relapse. In addition tofamily and peer support, there were participants who felt that spiritual support was important inhelping them cope with the possibility of relapse. The survivors felt that they did not receive enoughinformation about post-treatment care, dietary measures and supplements for relapse prevention,treatment and prognosis.Conclusion: Prostate cancer has a significant impact on the survivor’s lifestyle, emotional andphysical health. They need information and emotional support from the healthcare professionals,family and peers. Therefore, it is important for healthcare providers to explore the needs of prostatecancer survivors and provide the necessary support.

Juvenile recurrent respiratory papillomatosis: A rare masquerade of asthma

Juvenile recurrent respiratory papillomatosis (JRRP) is a rare condition. The varied presentation of this condition predisposes to misdiagnosis and potential life-threatening airway obstruction. In this paper, we have reported a case of JRRP presenting as severe respiratory distress and consequently mistreated as asthmatic attack culminating in a near fatal acute airway obstruction.

Malaysian private general practitioners’ views and experiences on continuous professional development: A qualitative study

Introduction: Continuous professional development (CPD) is an important aspect of a medical practitioner’s career. Aiming to be at par with other developed countries for high quality of professional practice, Malaysia is planning to implement compulsory CPD for the doctors. Aim: The aim of the study was to explore the private general practitioners’ (GPs) views, experiences and needs regarding CPD programme in the primary care service. Methods: This study used a qualitative methodology. Seven semi-structured interviews and three focus group discussions were conducted with private general practitioners from an urban area of Malaysia between January and December 2012. An interview topic guide was developed based on literature review and researchers’ discussions and it was used to guide the interviews. All the interviews were audio-recorded, transcribed verbatim and the transcripts formed the data for analysis using the thematic approach. Results: GPs undertook a wide range of CPD programmes to keep up with medical advances, meet patients’ expectations and improve financial rewards. Conferences, lectures and online recourses were the most mentioned methods of keeping updated. Some of the GPs felt that peer motivation and networking seem to motivate and facilitate participation in CPD programmes. However, they were wary of the validity and relevance of some CPD programmes, particularly those related to pharmaceutical industry. Although the participants agreed to the new mandatory CPD regulation, they voiced concerns on how it would be implemented and wished for a more effective method of monitoring. Conclusions: Organised peer support and relevant CPD content may improve GP participation in CPD but adequate regulatory measure should be in place to monitor the CPD activities.

“Personal mission statement”: An analysis of medical students’ and general practitioners’ reflections onpersonal beliefs, values and goals in life

Background: Personal mission in life can determine the motivation, happiness, career advancement and fulfilment in life of the medical students (MSs) along with improvement in professional/clinical performance of the family physicians. This study explored the personal beliefs, values and goals in the lives of MSs and general practitioners (GPs). Methods: Fourth-year MSs at the Universiti Putra Malaysia and GPs who participated in a 2-hour session on ‘Ethics in Family Medicine’ in 2012 were invited. All the participants submitted the post-session written reflections about their personal missions in life. The written reflections were analysed using thematic analysis. Results: A total of 87 MSs and 31 GPs submitted their written reflections. The authors identified 17 categories from the reflections contained by four themes—good vs. smart doctor, professional improvement vs. self-improvement, self-fulfilment and expressed motivation. The most common categories were “to be a good doctor” (97/330) and “professional improvement”(65/330). Many MSs had expressed motivation and wanted to be a smart doctor as compared to the GPs, whereas a larger number of GPs wished to have a fulfilled life and be a good doctor through professional improvement. Conclusion: The difference between the two student groups might indicate different levels of maturity and life experiences. Medical teachers should engage students more effectively in orientating them towards the essential values needed in medical practice.

The outcome of patients with renal involvement in pediatric-onset systemic lupus erythematosus--a 20-year experience in Asia.

Systemic lupus erythematosus (SLE) predominantly affects women of childbearing age, but 15-20% of cases are diagnosed during childhood. It is important for physicians to understand the epidemiology and clinical presentation for early detection and diagnosis of this disease in difference races. The aim of this retrospective review was to provide a 20-year experience for initial clinical and laboratory manifestations and outcomes in pediatric-onset SLE (pSLE) in a medical center in Asia. We reviewed medical records between April 1990 and June 2012 of patients with a diagnosis of International Classification of Diseases, Ninth Revision (ICD-9) code 710.0 (SLE), who admitted or received follow-up in the Department of Pediatrics at Chang Chung Memorial Hospital. Patients with a diagnosis of SLE prior to their 18th birthday and followed up at our hospital were eligible for inclusion in this study. Medical records regarding age, gender, date of birth and diagnosis, clinical manifestations at diagnosis, laboratory results, image studies and the classification criteria were reviewed. Patients received regular outpatient department follow-up and laboratory survey every 1-6 months. The study cohort consisted of 189 patients; 164 females (86.87%) and 25 males (13.23%). The overall mean age at pSLE diagnosis was 12.62 ± 2.77 years. The most common clinical symptom was malar rash, followed by arthritis and oral ulcers. There was no significant difference in clinical and laboratory manifestations between females and males. More than half of the patients presented with renal involvement initially. The most common histological finding was Class IV lupus nephritis (LN), especially in males (p = 0.034) and young age. Even with severe LN, the rate of end-stage renal disease (ESRD) was low if adequate treatment was initiated. The 5, 10 and 15-year ESRD-free survival rates were 95.4%, 94.0% and 89.9% in patients with biopsy-proven LN. However, infection was the leading cause of mortality. Therefore, aggressive treatment for major organ involvement is important, but physicians must also be aware of fatal infection. The overall survival rates were 5 years: 93.4% and 10-20 years: 89.6%.

Perception of Quality of Life among People with Diabetes.

Diabetes is a chronic disease that affects a patient's quality of life. This cross-sectional study aimed to determine the socio-demographic and disease profile factors associated with poor quality of life among patients with diabetes. The study was conducted at a primary health care clinic in Kuching between August to November 2010. Short Form - 36 (SF - 36) questionnaire was used to assess the quality of life of diabetic patients aged ≥ 18. A total of 142 respondents participated in the survey. After adjusting for age, those with no education scored lower at vitality (p=0.043) and emotional health (p=0.033) compared with those who have tertiary education. Those working in the private sector scored better for physical functioning (p=0.042) compared with pensioners and the unemployed. Patients with uncontrolled diabetes scored lower in the role-emotional domain (p=0.003). Participants who were on <3 (p=0.014) and ≥3 (p=0.024) oral medications had better score for role-physical than those on insulin. Those on insulin had worse score for bodily pain than those on oral medication only (vs <3 oral drugs, p=0.026; vs ≥3 oral drugs, p=0.001). Various socio-demographic factors, uncontrolled diabetes and insulin usage were found to have negative impact on a diabetic patient's quality of life. Programmes addressing the physical and emotional needs of diabetic patients at the primary health care setting are essential to help improve their quality of life.

Childhood Obesity, Self-Esteem and Health-Related Quality of Life among Urban Primary Schools Children in Kuching, Sarawak, Malaysia

Introduction: There is limited data on childhood obesity and its impact on children from diverse cultural backgrounds. This study is aimed at determining the association between obesity, self-esteem and health-related quality of life (HRQOL) among Malaysian urban primary school children of different ethnicity. Methods: A cross-sectional study was conducted involving 311 children aged 11-13 years from primary schools in Kuching, Sarawak. Self-esteem and healthrelated quality of life (HRQOL) were measured using the Lawrence Self-esteem Questionnaire (LAWSEQ) questionnaire and the Paediatric Quality of Life Inventory (PedsQL), respectively. Body weight and height were taken and body mass index for age calculated. Results: The prevalence of overweight and obesity among the children were 18.2% and 15.2% respectively. Parent-proxy and child self-reported PedsQL scores were higher for normal weight children compared to thin and obese children, but lower than overweight children. At the subscale level, only parent-proxy PedsQL scores in psychosocial health and emotional component were significantly different between overweight and obese children (p=0.019, p=0.02). The Self-esteem score was significantly correlated with parent and child PedsQL scores. Although obesity was associated with lower HRQOL among children, both parent and child PedsQL scores among the overweight group were higher than that for the normal weight group. Overweight and obesity did affect quality of life and self-esteem of children in this study, particularly in the areas of psychosocial and emotional health. Conclusion: Policy makers and programme managers should take into consideration the impact of obesity on children and parents in designing intervention programmes.

Perception of quality of life among people with diabetes

Diabetes is a chronic disease that affects a patient’s quality of life. This cross-sectional study aimed to determine the socio-demographic and disease profile factors associated with poor quality of life among patients with diabetes. The study was conducted at a primary health care clinic in Kuching between August to November 2010. Short Form - 36 (SF - 36) questionnaire was used to assess the quality of life of diabetic patients aged ≥ 18. A total of 142 respondents participated in the survey. After adjusting for age, those with no education scored lower at vitality (p=0.043) and emotional health (p=0.033) compared with those who have tertiary education. Those working in the private sector scored better for physical functioning (p=0.042) compared with pensioners and the unemployed. Patients with uncontrolled diabetes scored lower in the role-emotional domain (p=0.003). Participants who were on <3 (p=0.014) and ≥3 (p=0.024) oral medications had better score for role-physical than those on insulin. Those on insulin had worse score for bodily pain than those on oral medication only (vs <3 oral drugs, p=0.026; vs ≥3 oral drugs, p=0.001). Various socio-demographic factors, uncontrolled diabetes and insulin usage were found to have negative impact on a diabetic patient’s quality of life. Programmes addressing the physical and emotional needs of diabetic patients at the primary health care setting are essential to help improve their quality of life.

Risk factors for diabetic retinopathy in diabetics screened using fundus photography at a primary health care setting in East malaysia.

INTRODUCTION: This study reports on the prevalence of diabetic retinopathy (DR) and risk factors among diabetic patients, who underwent fundus photography screening in a primary care setting of Borneo Islands, East Malaysia. We aimed to explore the preliminary data to help in the planning of more effective preventive strategies of DR at the primary health care setting. MATERIALS AND METHODS: A cross-sectional study on 738 known diabetic patients aged 19-82 years was conducted in 2004. Eye examination consists of visual acuity testing followed by fundus photography for DR assessment. The fundus pictures were reviewed by a family physician and an ophthalmologist. Fundus photographs were graded as having no DR, NPDR, PDR and maculopathy. The data of other parameters was retrieved from patient's record. Bi-variate and multivariate analysis was used to elucidate the factors associated with DR. RESULTS: Any DR was detected in 23.7% (95% CI=21 to 27%) of the patients and 3.2% had proliferative DR. The risk factors associated with any DR was duration of DM (OR =2.5, CI=1.6 to 3.9 for duration of five to 10 years when compared to <5 years) and lower BMI (OR=1.8, CI=1.1 to 3.0). Moderate visual loss was associated with DR (OR=2.1, CI=1.2 to 3.7). CONCLUSIONS: This study confirms associations of DR with diabetic duration, body mass index and visual loss. Our data provide preliminary findings to help to improve the screening and preventive strategies of DR at the primary health care setting.

Brief report do university students have high cardiovascular risk? A pilot study from universiti malaysia sarawak (unimas).

A health screening was done in UNIMAS in August 2008 for 237 undergraduate students. Body mass index (BMI), waist circumference (WC) and blood pressure (BP) were measured for all subjects. Total cholesterol and glucose levels were checked for those who fulfilled the screening criteria. The proportion of participants with cardiovascular (CVD) risk factors was high. The strategies for health promotion should not only be targeted to the older community but also to the younger community.

Street ketamine-associated bladder dysfunction: an emerging health problem.

INTRODUCTION: Ketamine is frequently abused nowadays as a recreational drug. Case reports are emerging since 2007 to describe a new clinical entity of severe bladder dysfunction associated with chronic abuse of street ketamine. CLINICAL PRESENTATION: Severe lower urinary tract symptoms of urinary frequency and urgency which are refractory to conventional treatment. Quality of life is adversely affected as a consequence. Chronic kidney disease will develop in advanced cases. Investigation findings: The urine is sterile on culture. Ultrasound will show reduced bladder capacity with thickened bladder wall. In advanced stage, hydronephrosis and renal impairment will develop. TREATMENT: Patients should be advised to stop street ketamine use immediately. Anticholinergic medication could be tried to alleviate the symptoms. Refractory cases with dilatation of the upper urinary tract might need urinary diversion. CONCLUSION: Awareness of this new condition is essential in diagnosis. Early intervention offers better treatment outcome.

Penetration of topically applied levofloxacin into eyes with thin-wall filtering bleb after trabeculectomy.

PURPOSE: To investigate the comparative penetration of 0.3% levofloxacin eye drops into the aqueous humour between cataract patients with or without (control) thin-wall filtering blebs. METHODS: One drop of 0.3% levofloxacin was administered to the eyes at 30-min intervals for 3.5 h before phacoemulcification for both groups. Aqueous humour samples (0.1-0.2 ml) were aspirated during surgery. The concentration of levofloxacin in the aqueous humour was determined by high-performance liquid chromatography. The Student's t-test, Pearson correlation, and chi(2) test were used to compare the data of the two groups. A P<0.05 was required for results to be considered statistically significant. RESULTS: The levofloxacin concentration in the aqueous humour was significantly increased (P<0.0001) in the bleb (mean+SD: 3.7+/-2.3 microg/ml) vscontrol group (0.4+/-0.2 microg/ml). Intraocular pressure and the bleb area were not correlated with levofloxacin concentration. CONCLUSION: The presence of thin-wall filtering blebs increases intraocular penetration of topically administered levofloxacin.