Serological prevalence of FeLV and FIV in cats in Peninsular Malaysia.

Feline Leukemia (Fel.V) and Feline Immunodeficiency Virus (FIV) are the two feline retroviruses that were studied in 65 client-owned cats from eight veterinary centres in selected areas throughout Peninsular Malaysia during a 4-week period. Blood samples were collected for serological tests using SensPERT®FeLV Ag/FIV Ab test kits. Five of 65 cats (7.69%) tested positive for the FeLV antigen and 10 cats (21.54%) tested positive for FIV antibodies. Only one cat had a dual infection. Chi square analysis revealed a significant association (P<0.05) between the health status and lifestyle of the cat and FeLV-positive status. FeLV infections were more likely to occur in a pedigree, adult male cat while FIV was more likely to infect the adult, intact cat that is aggressive towards other cats

Molecular Detection, Phylogenetic Analysis, and Identification of Transcription Motifs in Feline Leukemia Virus from Naturally Infected Cats in Malaysia

A nested PCR assay was used to determine the viral RNA and proviral DNA status of naturally infected cats. Selected samples that were FeLV-positive by PCR were subjected to sequencing, phylogenetic analysis, and motifs search. Of the 39 samples that were positive for FeLV p27 antigen, 87.2% (34/39) were confirmed positive with nested PCR. FeLV proviral DNA was detected in 38 (97.3%) of p27-antigen negative samples. Malaysian FeLV isolates are found to be highly similar with a homology of 91% to 100%. Phylogenetic analysis revealed that Malaysian FeLV isolates divided into two clusters, with a majority (86.2%) sharing similarity with FeLV-K01803 and fewer isolates (13.8%) with FeLV-GM1 strain. Different enhancer motifs including NF-GMa, Krox-20/WT1I-del2, BAF1, AP-2, TBP, TFIIF-beta, TRF, and TFIID are found to occur either in single, duplicate, triplicate, or sets of 5 in different positions within the U3-LTR-gag region. The present result confirms the occurrence of FeLV viral RNA and provirus DNA in naturally infected cats. Malaysian FeLV isolates are highly similar, and a majority of them are closely related to a UK isolate. This study provides the first molecular based information on FeLV in Malaysia. Additionally, different enhancer motifs likely associated with FeLV related pathogenesis have been identified

Synthesis and characterization of chitosan-saponin nanoparticle for application in plasmid DNA delivery

Nonviral delivery system receives attention over the last decade. Chitosan (CS) is a cationic polymer whereas saponin (SP) is classified as glycoside. In this study, a spherically-shaped CS-SP nanoparticle was synthesized and characterized. The ability of the nanoparticle to protect DNA from enzymatic degradation, its thermostability and cytotoxicity were evaluated. The particle size was found below 100 nm as determined by Zetasizer, transmission electron microscopy (TEM), and field scanning electron microscopy (FSEM) results. The surface charge ranges from 43.7 mV to 38.5 mV before and after encapsulation with DNA plasmid, respectively. In terms of thermostability, Thermal Gravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC) revealed that CS-SP nanoparticle had a melting temperature of 110°C, with rapid decomposition occurring at 120°C. Encapsulation of DNA with the synthesized nanoparticle was evidenced by changes in the FTIR spectra including characteristic peaks at 3267.39 and 1635.58 cm−1, wavenumbers. Additional peak was also observed at 1169.7 cm−1 following encapsulation. Electrophoretic mobility showed that CS-SP nanoparticle protected plasmid DNA from enzymatic degradation, while cell viability assays confirmed that the synthesized nanoparticle exhibited low cytotoxicity at different concentrations in avian cells. Taken together these, CS-SP nanoparticle showed potentials for applications as a DNA delivery system

Prevalence and risk factors of feline leukaemia virus and feline immunodeficiency virus in peninsular Malaysia

<p>Abstract</p> <p>Background</p> <p>Feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) are major causes of morbidity and mortality in domestic and wild felids. Despite the clinical importance of feline retroviruses and the growing interest in cats as pets, information about FeLV and FIV in Malaysia is presently insufficient to properly advise veterinarians and pet owners. A cross-sectional study was carried out from January 2010 to December 2010 to determine the prevalence and risk factors associated with FeLV and FIV among domestic cats in peninsular Malaysia. Plasma samples were harvested from the blood of 368 domestic cats and screened for evidence of FeLV p27 antigen and FIV antibodies, using an immunochromatographic kit. Additionally, data on cat demographics and health were collected using a structured questionnaire, and were evaluated as potential risk factors for FeLV or FIV status.</p> <p>Results</p> <p>Of the 368 cats that were evaluated in this study, 12.2% (45/368; 95% CI = 8.88 - 15.58) were positive for FeLV p27 antigen, 31.3%, (115/368; 95% CI = 26.51 - 35.99) were seropositive to FIV antibodies, and 4.3% (16/368; 95% CI = 2.27 - 6.43) had evidence of both viruses. Factors found to significantly increase the risk for FeLV seropositivity include sex, age, behaviour, sickness, and living in a multi-cat household. Seropositive response to FIV was significantly associated with sex, neuter status, age, behaviour, and health status.</p> <p>Conclusions</p> <p>The present study indicates that FeLV and FIV are common among domestic cats in peninsular Malaysia, and that factors related to cat demographics and health such as age, sex, behaviour, health status and type of household are important predictors for seropositive status to FeLV or FIV in peninsular Malaysia. High prevalence of FeLV or FIV observed in our study is of concern, in view of the immunosuppressive potentials of the two pathogens. Specific measures for control and prevention such as screening and routine vaccination are needed to ensure that FeLV and FIV are controlled in the cat population of peninsular Malaysia.</p

Molecular and pathological identification of feline coronavirus type I

The coronavirus in cats has been described as feline infectious peritonitis (FIPV) and feline enteric coronavirus (FECV). FIPV is highly fatal and caused immune-mediated pyogranulomatous disease, whereas FECV causes mild enteric infection. In this study, we described the isolation and molecular characterization of naturally occurring feline coronavirus from domestic cat in Malaysia. Additionally, the resultant pathological conditions observed in the infected cat were reported. Ascitic fluid sample was collected from 3-year-old domestic cat clinically suspected of effusive (wet form) FIP and subjected to virus isolation in Felis catus whole fetus cell cultures (Fcwf-4). The result of virus isolation was confirmed using one step reverse transcription polymerase chain reaction (RT-PCR). To gain insight on the genetic variant of FCoV, the S-gene sequence was amplified using type 1 specific primers. Virus isolation showed cytopathic effect (CPE) characterized by giant cells, ballooning and cells detachment. Ultrastructural findings showed virus particles attached to plasma membrane and later invaginated from the cell membrane. Virus-like particles were also observed in the vacuoles, likely as a result of spillage of mature virus-like particles into the cytoplasmic matrix. Histopathological examination of kidney, spleen and intestine organ samples revealed coagulative necrosis with infiltration of neutrophils and mononuclear cells. In conclusion, this study reports the first isolation of feline coronavirus in Malaysian cats and importantly the isolated virus was confirmed to be type I using S-gene amplification.Key words: Feline coronavirus, Fcwf-4, RT-PCR, effusive FIP, ultrastructure, histopathology

Investigations for the possible use of a monoclonal antibody produced against strongyloides ratti antigen as an immunodiagnostic reagent for active strongyloidiasis

Background: Currently, most of the available serological diagnostic kits for strongyloidiasis are based on the use of the crude antigens of Strongyloides ratti, which are good, but with less sensitivity towards the infection. Hence, this study aimed to produce and evaluate monoclonal antibody for detecting soluble parasite antigen in animal sera. Methods: The study was conducted in the Department of Medical Microbiology and Parasitology, University Putra Malaysia in 2014-2017. Saline extract protein from the infective larvae of S. ratti was used to immunize BALB/c mice and subsequent fusion of the B-cells with myeloma cells (SP2/0) using 50% PEG. The hybridomas were cultured in HAT medium and cloned by limiting dilutions. Positive hybrids were screened by indirect ELISA. The ascites fluid from the antibody-secreting hybridoma was purified and the MAb was characterized by western-blots and evaluated in sandwich ELISA for reactivity against the homologous and heterologous antigens. Results: An IgG1 that recognizes a 30 and 34 kDa protein bands was obtained. The MAb was recognized by all S. ratti-related antigens and cross-reacted with only Toxocara canis antigens in both assays. The minimum antigen detection limit was found to be 5 ng/ml. All antibody-positive rat and dog sera evaluated have shown antigen-positive reactions in Sandwich-ELISA. Conclusion: The MAb produced, was able to detect antigens in strongyloidiasis and toxocariasis in animal models and may also be useful for the serological detection of active strongyloidiasis and visceral toxocariasis in human sera

In vitro evaluation of antiviral properties of edible bird nest extract against feline infectious peritonitis virus

An in vitro study was carried out to evaluate the antiviral properties of edible bird nest extract (BNE) against feline infectious peritonitis virus (FIPV). Cytotoxicity assay was conducted towards BNE test material in Crandell Feline Kidney (CrFK) cells using MTT assay to determine the 50% cytotoxic concentration (CC50)values. For antiviral test, three treatments were used to determine the antiviral inhibition effect by BNE extract. Co-treatment [(V+E) +C] was done by mixing the virus(V) and extract(E) together before inoculating into cells(C). Pre-treatment [(E+C) +V], involved treatment of extract before inoculation of the cells with virus. Post-treatment [(V+C) +E] was done by inoculating the virus first into the cells before inoculation of extract. Ten-fold dilutions of BNE were used to determine the CC50 until 8th doubling dilutions. The FIPV dose was fixed for 100TCID50. Cytotoxicity assay showed that all concentrations could be used for antiviral assay except for the stock solution. The results also showed that the extract was non-toxic to the cells. For antiviral assay, all treatments showed inhibitory effects on virus multiplication in the cells where pre-treatment showed the highest effect compared to the other two treatments. However, the finding was not statistically significant from the control treatment groups (P<0.05)

First molecular detection of porcine bocavirus in Malaysia

Several strains of porcine bocaviruses have been reported worldwide since their first detection in Sweden in 2009. Subsequently, the virus has been reported to be associated with gastrointestinal and respiratory signs in weaner and grower pigs. Although Malaysia is host to a self-sufficient swine livestock industry, there is no study that describes porcine bocavirus in the country. This report is the first to describe porcine bocavirus (PBoV) in Malaysian swine herds. PBoV was identified in various tissues from sick and runt pigs using the conventional PCR method with primers targeting conserved regions encoding for the nonstructural protein (NS1) gene. Out of 103 samples tested from 17 pigs, 32 samples from 15 pigs were positive for porcine bocavirus. In addition, a higher detection rate was identified from mesenteric lymph nodes (52.9%), followed by tonsil (37.0%), and lungs (33.3%). Pairwise comparison and phylogenetic analyses based on a 658-bp fragment of NS1 gene revealed that the Malaysian PBoV strains are highly similar to PBoV3 isolated in Minnesota, USA. The presence of porcine bocavirus in Malaysia and their phylogenetic bond was marked for the first time by this study. Further studies will establish the molecular epidemiology of PBoV in Malaysia and clarify pathogenicity of the local isolates

Breaking the cycle of the COVID-19 transmission: a challenge for Nigeria

COVID-19 has already spread to almost every country in the world, including the arctic. The impact on human health has been severe, with an increasing number of fatalities. With the spread comes economic hardship due to the preventive strategies adopted. Movement restrictions imposed in Nigeria as a result of the outbreak have generated controversies among the poor masses that depend on daily hustles to fend for themselves and their families. Nigeria being a very populous country with a high number of low-income earners who depend on their daily efforts to get food for their families, control measures like the movement restrictions and closure of business premises would have a devastating impact on them. Although the government has responded with some palliative measures, it is evident that these interventions may not be sufficient, mainly due to potential malpractices that will end up denying many supposed beneficiaries