Leptospirosis, also known as “rat-urine disease”, is a neglected zoonotic and waterborne disease that is caused by Leptospira spp. This disease is transmitted by direct and indirect exposure to the urine and stool of infected animals. The current estimate has highlighted that leptospirosis has caused at least one million cases and 60,000 deaths, with high endemicity in tropical regions. With climate change, urbanisation, and increasing human-animal interaction, the threat of leptospirosis and other zoonotic diseases will continue to emerge. Investing in multidisciplinary research, technology, and global collaboration is critical to anticipate, detect, and respond effectively to these evolving threats.

The genus Vibrio forms associations with phytoplankton, using algal cells as vectors for proliferation and survival in aquatic environments. These interactions serve as a reservoir for Vibrio species, influencing the ecology of estuarine and potentially affecting the concentration and distribution of pathogenic Vibrio. Despite extensive research on Vibrio and phytoplankton interactions, little is known about their associations with key environmental factors such as temperature, pH, and salinity. This study aimed to determine the concentrations of Vibrio species, environmental factors influencing their proliferation, and analyse Vibrio-phytoplankton interactions. Water samples were collected from two estuaries, with physicochemical parameters recorded in-situ. Phytoplankton abundance was determined microscopically, while Vibrio isolation was conducted using TCBS agar. Molecular identification of Vibrio species was performed using PCR amplification of species-specific genes (OmpW for V. cholerae and ToxR for V. parahaemolyticus). The results showed that 36.7% (n=22/60) of the samples positive for V. cholerae and 31.7% (n=19/60) were positive for V. parahaemolyticus. None of the isolates tested positive for the ctxA and tdh virulence genes. Statistical analysis revealed significant differences (p < 0.05) between Vibrio concentrations, water temperature and salinity, but pH showed no significant effect. Pearson's correlation analysis revealed positive correlations between Vibrio concentrations with high temperature (31.2 – 34.5 ºC) and salinity (28.7 - 31.0 ppt), while exhibiting a negative correlation with phytoplankton concentration. Phytoplankton was influenced by temperature and salinity, with high salinity reducing abundance. The results of this study may provide insights into potential public health risks posed by Vibrio species in estuarine ecosystems and contribute to understanding their ecological dynamics in Northern Sarawak, Malaysia.

Over the past decade, the occurrence of milk-borne infections caused by Shiga toxin-producing Escherichia coli (STEC) and Salmonella enterica serovar Typhimurium (S. Typhimurium) has adversely affected consumer health and the milk industry. We aimed to detect and genotype the strains of E. coli and S. Typhimurium isolated from cow and goat milks using two genotyping tools, BOX-PCR and ERIC-PCR. A total of 200 cow and goat milk samples were collected from the dairy farms in First, E. coli and Salmonella spp. detected in the samples were characterized using PCRs to identify pathogenic strains, STEC and S. Typhimurium. Next, the bacterial strains were genotyped using ERIC-PCR and BOX-PCR to determine their genetic relatedness. Out of 200 raw milk samples, 46.5% tested positive for non-STEC, 39.5% showed the presence of S. Typhimurium, and 11% were positive for STEC. The two genotyping tools showed different discrimination indexes, with BOX-PCR exhibiting a higher index mean (0.991) compared to ERIC-PCR (0.937). This suggested that BOX-PCR had better discriminatory power for genotyping the bacteria. Our study provides information on the safety of milk sourced from dairy farms, underscoring the importance of regular inspections and surveillance at the farm level to minimize the risk of E. coli and Salmonella outbreaks from milk consumption.

Aquaculture activities in Sarawak is currently undergoing great expansion and advancement by the government to provide sustainable food source for the local people. With the changing environment, climate change and increases in nutrients this could disrupt the water quality for the fish production. Hence, this study was conducted to assess the water quality in commercialize aquaculture ponds in Southern Sarawak. This study was carried out between June 2023 to November 2023. The temperature, dissolved oxygen, turbidity, pH, conductivity, total dissolved solids, ammonia, phosphate, nitrate, and nitrite which are deemed as the most sensitive and pertinent for the fish production were evaluated. The YSI multiparameter was used to measure the physical variables and the nutrients variables were measured using the HACH spectrophotometer. The results obtained for the temperature (28±0.00-32±0.00 °C), dissolved oxygen (1.10±0.01-12.99±0.01 mg/L), pH (3.72±0.03-8.39±0.06 mg/L), conductivity (0.03±0.00- 215.10±0.35 μS/cm), total dissolved solids (10.45±0.01-10137.17±0.02 mg/L), turbidity (6.77±0.16-483.22±40.41 NTU), ammonia (0-3.5 mg/L), phosphate (0.06±0.03-4.52±0.04 mg/L), nitrate (0.01±0.00-3.5 mg/L) and nitrite (0.003±0.002-3.5 mg/L) denoted that the physicochemical parameters of water quality in the aquaculture ponds, if found within normal levels considered tolerable. Some of the physicochemical parameters examined this study did not fall within the recommended range for optimal fish production, suggesting that some physicochemical parameters in these aquaculture ponds does not provide conducive conditions for the fish survival and growth. Thus, vigilant monitoring and stricter management of water quality in aquaculture ponds is needed to protect the health of aquaculture industry as well as the public health that consume them.

Leptospirosis is a severe and potentially fatal re-emerging zoonotic and waterborne disease caused by pathogenic and intermediate species of Leptospira. Given the high global rates of morbidity and mortality associated with this disease, there is an urgent need to explore alternative therapeutic agents to enhance treatment options. This study investigates the anti-leptospiral efficacy of several common antibiotics-penicillin G, doxycycline, ampicillin, amoxicillin, cefotaxime, chloramphenicol, and erythromycin, as well as extracts from local herbs, Hydnophytum formicarum Jack and Boesenbergia stenophylla, against pathogenic and intermediate Leptospira strains. A broth microdilution method determined the minimum inhibitory concentration (MIC) for the antibiotics and herb extracts. Both herbs were extracted using four different solvents: ethyl acetate, methanol, hexane, and chloroform. The extracts were then analysed using gas chromatography-mass spectrometry (GC-MS) to identify their phytochemical compounds. The results demonstrated that cefotaxime and erythromycin exhibited the highest anti-leptospiral activity, with MIC values of 0.2 µg/mL. This was followed by amoxicillin and ampicillin (0.2-0.39 µg/mL), penicillin G (0.39-3.13 µg/mL), chloramphenicol (0.78-3.13 µg/mL), and doxycycline (0.78-12.5 µg/mL). H. formicarum Jack and B. stenophylla extract extractions displayed the lowest MICs (62.5 µg/mL) for the ethyl acetate, methanol, and hexane extracts. They contained various phytochemical constituents, including some with anti-leptospiral properties. These findings indicate that different strains of Leptospira respond with varying levels of inhibition to the antibiotics and herb extracts studied. The extracts from H. formicarum Jack and B. stenophylla may have potential as anti-leptospiral drugs. However, further in-vivo studies are needed to better understand their efficacy against Leptospira.

This study is to analyse the spatial distribution of leptospirosis and identify its high and low incidence clusters in Sarawak. The annual incidence rate at the district level was calculated using confirmed report of leptospirosis cases from year 2011 to 2018. Empirical Bayes estimation smoothing of relative risks was used to display the spatial distribution of leptospirosis across the study region. Moran's Global Index and Local Indicators of Spatial Association (LISA) were used to analyse the existence of global and local spatial autocorrelation. Data were analysed using ArcGIS and Geoda software at the district level. The annualised average incidence reported during the study period was 20.83 per 100,000 population, with the highest cases reported in year 2018 (n = 870). The Global Moran's Index revealed spatial clustering of leptospirosis incidence in 2012 (Moran's I: 0.23), 2013 (Moran's I: 0.33), and 2014 (Moran's I: 0.45), while 2011 (Moran's I: -0.01), 2015 and 2016 (Moran's I: 0.09), 2017 (Moran's I: 0.13), and 2018 (Moran's I: 0.04) showed random patterns. High incidence clusters of leptospirosis were primarily congested in the Southeast of Sarawak, involving districts such as Kapit, Belaga, Song, Tatau and Lubok Antu. Spatial and temporal patterns of leptospirosis incidence were heterogeneous across Sarawak. This study facilitates the implementation of targeted interventions and control measures for leptospirosis in Sarawak by identifying spatial cluster and outliers.

Waterborne parasites, particularly Cryptosporidium and Giardia, are emerging pathogens implicating the safety level of drinking water globally. The aim of this study was to determine the distribution pattern of waterborne parasites in raw and treated water at urban and rural water treatment plants and untreated water from gravity-feed system in Kuching, Sarawak. This study focused on water treatment plants (four urban and two rural) and Bong rural community that utilise gravity-feed system in Kuching, Sarawak. A total of 69 raw and treated water samples were collected and processed before being used in detection of Cryptosporidium and Giardia using Aqua-Glo™ G/C Direct and 4′,6-diamidino-2-phenylindole stains, as well as other parasites that were detected using Lugol’s iodine staining. Parameters which were temperature, pH, turbidity, dissolved oxygen, total dissolved solids, conductivity, faecal coliform of the water as well as rainfall intensity were determined. Correlation of the parameters with distribution of the waterborne parasites was analysed. Out of 69 water samples collected across all localities, 25 samples were contaminated with waterborne parasites with varying waterborne parasite concentration in the water samples. The presence of waterborne parasites in the raw and treated water of water treatment plants in this study signifies public health threats do exist despite being conventionally treated. This study also highlights that the gravity-feed system which is commonly depended by rural communities in Malaysia may facilitate waterborne parasitic infections.

Aims: The gut microbiota is referred to as an 'extra organ' and is critical in assisting the host in terms of nutrition and immunity. Environmental stressors could alter the gut microbial community and cause gut inflammation. This study aimed to investigate and compare the gut microbiota community between healthy and diseased Tor tambroides. Methodology and results: In this study, such gut microbial alterations were explored using NGS-based 16S rDNA targeted sequencing on the Malaysian mahseer (T. tambroides). Three healthy adult and three diseased adult Malaysian mahseers (showing signs of exophthalmia, coelomic distension and petechial haemorrhage) were obtained from LTT Aquaculture Sdn Bhd. Our results revealed significant differences in microbial diversity, composition and function between both populations of T. tambroides. Alpha diversity analysis depicts lower diversity of gut microbiota composition in diseased T. tambroides as compared to the healthy group. In particular, Enterobacteriaceae, Aeromonas, Bacteroides, Vibrio and Pseudomonas were found within gut microbiota of the diseased fishes. In addition, cellulose degrading bacteria and protease-producing bacteria were identified from the gut of T. tambroides. Conclusion, significance and impact of study: Thus, our findings emphasized on the association between the alteration in gut microbiota composition and infectious abdominal dropsy (IAD) in T. tambroides. This finding is important to provide basic information for further diagnosis, prevention and treatment of intestinal diseases in fish.

The spatial distribution of environmental conditions may influence the dynamics of vectorborne diseases like leptospirosis. This study aims to investigate the global and localised relationships between leptospirosis with selected environmental variables. The association between environmental variables and the spatial density of geocoded leptospirosis cases was determined using global Poisson regression (GPR) and geographically weighted Poisson regression (GWPR). A higher prevalence of leptospirosis was detected in areas with higher water vapour pressure (exp(â): 1.12; 95% CI: 1.02 - 1.25) and annual precipitation (exp(â): 1.15; 95% CI: 1.02 - 1.31), with lower precipitation in the driest month (exp(â): 0.85; 95% CI: 0.75 - 0.96) and the wettest quarter (exp(â): 0.88; 95% CI: 0.77 - 1.00). Water vapor pressure (WVP) varied the most in the hotspot regions with a standard deviation of 0.62 (LQ: 0.15; UQ; 0.99) while the least variation was observed in annual precipitation (ANNP) with a standard deviation of 0.14 (LQ: 0.11; UQ; 0.30). The reduction in AICc value from 519.73 to 443.49 indicates that the GWPR model is able to identify the spatially varying correlation between leptospirosis and selected environmental variables. The results of the localised relationships in this study could be used to formulate spatially targeted interventions. This would be particularly useful in localities with a strong environmental or socio-demographical determinants for the transmission of leptospirosis.

Plant growth promoting rhizobacteria (PGPR) are strains of naturally occurring soil bacteria that live in close vicinity to the plant’s rhizosphere region which possess the capability to augment host growth. This study was conducted to isolate and identify potential PGPR isolates indigenous to Metroxylon sagu, Rottb. rhizosphere. These potential isolates were characterised based on their beneficial plant growth promoting (PGP) properties and identified by molecular analysis via 16S rDNA sequencing. A total of 18 isolates were successfully isolated, out of which five isolates were tested, and designated as (S1A, S2B, S3A, S3C and S42). Among the five isolates, two isolates (S2B and S3C) were found to produce high levels of indole-3-acetic acid (2.96 μg/mL and 10.31 μg/mL), able to fix nitrogen and show significant activity in phosphate solubilisation. The analysis of their sequences via National Center for Biotechnology Information (NCBI) suggested their close identity towards Lysinibacillus sphaericus and Bacillus thuringiensis. It can be concluded that the isolated PGPR possesses beneficial PGP attributes. It can be implied that the isolated PGPR are potential to be used as inoculant biofertilisers, beneficial for Metroxylon sagu, Rottb. growth. Hence, further studies need to be done to evaluate the effectiveness of the beneficial microbes towards sago seedlings growth, under pot experiment.

Background: Contaminated vegetables with intestinal parasites, particularly those eaten raw, represent a proportion of risks for humans acquiring foodborne parasitic diseases worldwide.Unfortunately, the risk is often neglected as limited studies have been reported about the parasitic occurrence from vegetables retailed in Malaysia.Objective: This study was conducted to determine the occurrence of intestinal parasites in fresh retailed vegetables in Kuching and Kota Samarahan, Sarawak.Methods: A total of 108 vegetables (comprising leafy and root-type vegetables) were purchased from three supermarkets and three wet markets in Kuching and Kota Samarahan.The samples were processed with 0.95% sodium chloride solution and underlaid with Sheather's sucrose solution.Cryptosporidium and Giardia were detected using AquaGlo TM G/C antibody reagent and 4′,6-diamidino-2-phenylindole stain.Other parasites were detected using Lugol's iodine stain.Results: A total of 24 out of 108 vegetables samples (22.2%) were contaminated with nematode larvae (range: 0.01 -0.71 larvae/g), Cryptosporidium oocysts (range: 0.01 -0.03 oocysts/g), hookworm ova (range: 0.01 -0.02 ova/g) and Giardia cyst (0.01 cysts/g).There were no significant differences (p > 0.05) for means of parasite concentrations in vegetables from supermarkets and wet markets, as well as parasite concentrations from leafy-type and root-type vegetables. Conclusion:The findings revealed relatively high numbers of intestinal parasites detected from fresh retailed vegetables, signifying potential foodborne transmission of parasitic infections if the vegetables are not prepared hygienically or cooked well.Besides, a high proportion of such infection risk may be reduced if farmers also take part in proper sanitation practices during vegetable production and transportation.

Intestinal parasitic infections are endemic in rural settings and may account for asymptomatic infections to various health complications. These infections are a cause of concern for communities of lower economic status, especially in developing countries. In Sarawak, indigenous populations residing in geographically inaccessible areas are socially and economically disadvantaged. Through close association with nature, these populations are prone to intestinal parasitism. Currently, scattered information has led to a continual state of neglect at each level of parasitic infection control. This urges for a review of their distribution and transmission based on previous reports to understand the pattern of the diseases in the state which can further address the improvement of mass controlling programs. A literature search was conducted to collect previous reports on human intestinal parasites in Sarawak, East Malaysia from PubMed (Medline), SCOPUS, ScienceDirect and Web of Science from January 2019 to March 2021. Extrapolating the current data in Sarawak which is still considered limited, further interdisciplinary strategies are demanded to give insights in the epidemiology and true prevalence of intestinal parasites in Sarawak. This review addresses for redirection of attitude towards intestinal parasitic infections where it should be given with ample attention by rural populations. In tandem to that, improvement of rural livelihood such as standard of living and sanitation in Sarawak should be accredited as part of the efforts to reduce the number of intestinal parasitic infections in the state. As a control measure, mass deworming should be reconsidered especially to the rural populations.

Through the advancement of biotechnology, DNA-based methods are the most effective techniques in species identification, as they are rapid and have higher stability in harsh conditions compared to protein-based methods. This study was conducted to determine the efficiency of the traditional DNA extraction method, phenol/chloroform/isoamyl alcohol (PCIA), and comparing it with the commercially available kit by evaluating the purity, concentration, and suitability for amplification of porcine DNA in raw chicken and beef mixtures. The quantity and quality of the DNA extracts were assessed using a UV-Vis spectrophotometer. Polymerase chain reaction (PCR) was performed using species-specific primers targeting mitochondrial DNA cytochrome b (cyt b) gene of chicken (227-bp), beef (274-bp), and pork (398-bp), to confirm the template usability and quality of the DNA extracts. High DNA concentrations and purity were obtained from meat samples extracted using the PCIA method. The visualization of pork DNA on 2% agarose gel was able to detect pork contamination in raw meat mixtures up to minute proportion (1%). The existence of pork in chicken and beef was indicated with the presence of a specific 398-bp DNA band. Thus, the PCIA method can be recommended as a cost-effective and an excellent alternative to more expensive extraction kits in detecting pork DNA in raw meat mixtures.

Aim This work reports a new method for the use of lasers for the selective killing of bacteria targeted using light-absorbing Silver nanoparticles (Ag-NPs) conjugated with a specific antibody against the Gram-positive bacterium Staphylococcus aureus (S. aureus). Methods and Results Ag-NPs were synthesized using a chemical reduction method and characterized with respect to their surface plasmon resonance, surface morphology via transmission electron microscopy (TEM) and dynamic light scattering (DLS). The bacterial surface was targeted using 20 nm Ag-NPs conjugated with an anti-protein A antibody. Labelled bacteria were irradiated with blue visible laser at 2 center dot 04 W/cm(2). The antibacterial activity of functionalized Ag-NPs was investigated by fluorescence microscopy after irradiation, and morphological changes in S. aureus after laser treatment were assessed using scanning electron microscopy (SEM). The laser-irradiated, functionalized Ag-NPs exhibited significant bactericidal activity, and laser-induced bacterial damage was observed after 10 min of laser irradiation against S. aureus. The fluorescence microscopic analysis results supported that bacterial cell death occurred in the presence of the functionalized Ag-NPs. Conclusions The results of this study suggest that a novel method for the preparation of functionalized nanoparticles has potential as a potent antibacterial agent for the selective killing of resistant disease-causing bacteria. Significance and Impact of the Study This study shows that Ag-NPs functionalized with a specific antibody, could be used in combination with laser radiation as a novel treatment to target resistant bacterial and fungal pathogens with minimal impact on normal microflora.

Background Leptospirosis is a zoonotic disease caused by spirochete bacteria in the genus Leptospira, and it has become a significant public health challenge in Malaysia. Environmental survival and persistence of this pathogen are highly dependent on environmental conditions such as moisture content, pH and temperature. These conditions are further adapted by the natural climate system including precipitation and humidity which is highly heterogeneous at a geographical scale. This paper described the spatial and temporal distribution of leptospirosis incidence with climate factors using Geographical Information System and stratified the climate factors based on their association with the disease incidence rate. Methods Leptospirosis surveillance data from 2012-2016 were integrated into this study along with seven geo-spatial climate variables for the state of Sarawak, Malaysia. High and low clustering of incidence rate was explored by Getis Ord Gi* statistics. Geographical Weighted Regression model was utilized to study the relationship between the incidence rate and selected climate variables. Results Spatial analysis revealed seven districts in the state of Sarawak as hot spot areas, and six cold spot areas with GiZ score varies between (-3.092 to 3.203). The cumulative incidence rate demonstrated an increasing trend towards the South-East region of Sarawak with an average of 162 cases per 100,000 population. The univariate analysis reported a significant relationship (p<0.05) between leptospirosis incidence rate and temperature seasonality with the lowest AIC value of 31741.4. The results showed that temperature seasonality explained 99% (R2: 0.99) of the spatial variances in incidence rate with 42.31% of the localities showing a significant positive relationship. Conclusion The present study highlighted the importance of temperature seasonality as a potential determinant for the spatial distribution of leptospirosis cases with different strength and direction of the association. In conclusion, this suggests that specific interventions at a locality with strong climate determinants should be implemented to combat the burden of leptospirosis.

Many cholera outbreaks worldwide were associated with cholera toxin-producing Vibrio cholerae. The bacteria are ubiquitous in aquatic environment, whilst phytoplankton is associated with adaptation of the Vibrio species. This study was conducted to detect cholera toxin-producing Vibrio cholerae, and to determine association of the selected water physicochemical parameters with the number of the bacteria. In this study, a total of ten phytoplankton samples were collected at Santubong and Samariang Estuaries in Kuching, Sarawak. Water physicochemical parameters (temperature, pH and salinity) were recorded. Vibrio bacteria were cultivated on thiosulfate citrate bile-salts sucrose selective agar and analysed for cholera toxin-producing Vibrio cholerae using polymerase chain reaction by targeting ctxA gene that encodes for virulence cholera enterotoxin subunit A. The result revealed that a range of 1.0 × 107 – 8.0 × 107 CFU/ml of yellow colonies growing on the thiosulfate citrate bile-salts sucrose agars. Inversely, no samples were positive with cholera toxin-producing Vibrio cholerae. The physicochemical parameters at Samariang Estuary were more associated with the number of bacteria in the samples compared to Santubong Estuary.

Silver nanoparticles (Ag-NPs) possess excellent antibacterial properties and are considered to be an alternative material for treating antibiotic-resistant bacteria. The present study was aimed at enhancing the antibacterial efficiency of Ag-NPs using visible laser light against Escherichia coli and Staphylococcus aureus in vitro . Four concentrations of Ag-NPs (12.5, 25, 50, and 100 μ g/ml), synthesized by the chemical reduction method, were utilized to conduct the antibacterial activity of prepared Ag-NPs. The antibacterial efficiencies of photoactivated Ag-NPs against both bacteria were determined by survival assay after exposure to laser irradiation. The mechanism of interactions between Ag-NPs and the bacterial cell membranes was then evaluated via scanning electron microscopy (SEM) and reactive oxygen species analysis to study the cytotoxic action of photoactivated Ag-NPs against both bacterial species. Results showed that the laser-activated Ag-NP treatment reduced the surviving population to 14% of the control in the E. coli population, while the survival in the S. aureus population was reduced to 28% of the control upon 10 min exposure time at the concentration of 50 μ g/ml. However, S. aureus showed lower sensitivity after photoactivation compared to E. coli . Moreover, the effects depended on the concentration of Ag-NPs and exposure time to laser light. SEM images of treated bacterial cells indicated that substantial morphological changes occurred in cell membranes after treatment. The results suggested that Ag-NPs in the presence of visible light exhibit strong antibacterial activity which could be used to inactivate harmful and pathogenic microorganisms.

Background and Objectives: Leptospirosis is a death-causing disease caused by corkscrew-shaped bacteria, Leptospira especially in tropical countries.Current study was aimed to detect pathogenic, intermediate and saprophytic Leptospira species using polymerase chain reaction (PCR) assay from an oil palm plantation in Borneo, specifically in Miri, Malaysia.Materials and Methods: A total of 63 samples from rodents (n = 3), water (n = 30) and soil (n = 30) were isolated from an oil palm estate in Northern Sarawak, Borneo.All samples were inoculated into modified semisolid Ellinghausen-McCullough-Johnson-Harris (EMJH) broth with 5-fluorouracil and incubated for a month.Polymerase chain reaction (PCR) was performed using primer targeting lipL32 (423 bp) for pathogenic, 16S rRNA (331 bp) for intermediate and rrs (240 bp) for saprophytic species.Results: pathogenic Leptospira was found in 33.3% rodents (1/3) Rattus tiomanicus, 23.3% soil samples (7/30) and 16.7% water samples (5/30).Intermediate species were demonstrated in the other 66.7% rodents (2/3), Sundamys muelleri and Rattus exulans and 10% soil samples (3/30).Saprophytic species was found in only 3.3% soil sample (1/30).Results from DNA sequencing analysis indicated that the most dominant pathogenic Leptospira species discovered in the study was Leptospira interrogans, followed by Leptospira noguchii and Leptospira weilii.Conclusion: These preliminary findings provide baseline data on the occurrence of Leptospira species in captured rodents and the environment.These findings could assist in control and prevention of leptospirosis among oil palm estate workers in Sarawak.Awareness and knowledge on leptospirosis should be promoted among oil palm workers for prevention and mitigation.

Aims: Leptospira spp. has the ability to develop biofilm communities and this attribute is an essential factor to leptospiral pathogenesis. This study aims to assess and quantify the biofilm forming ability of intermediate and saprophytic Leptospira strains. Methodology and results: The biofilm assay was quantified on microtitre polystyrene plates (abiotic) and wood chips (Jelutong Paya hardwood) over a duration of 11 days. Phase contrast light microscope was used to assess the structure of the on the surface. The biofilm production on wood chips surface were approximately one times higher than on polystyrene plate surface indicating Leptospira strains were capable of forming higher quantity of biofilm on biotic surface compared to abiotic surface by both intermediate and saprophytic Leptospira. A significant difference (p<0.05) exists in biofilms produced by Leptospira on wood surface which formed more biofilm than on polystyrene surface. The strongest biofilm producer is intermediate strain G14 with OD600 of 2.283 +/- 0.180 and OD600 of 2.333 +/- 0.037, on polystyrene and wood surface, respectively. Visualisation of biofilm by phase-contrast microscopy of two representative strains correlated with the OD values and the colour intensity of stained microtitre plates and wood surfaces. The biofilm formed comprises of a three-step process are adherence (1th to 24th h), maturation (6th to 7th day) and detachment (9th to 11th day) of biofilms. Conclusion, significance and impact of study: The contact time of intermediate pathogenic strains was faster compared to saprophytic strain, indicating the biofilm forming ability is related to the level of pathogenicity of Leptospira strains.

Listeria monocytogenes is one of the commonly isolated foodborne pathogens which cause illness, and listeriosis is a disease caused by this pathogen in human beings. Pets that consume contaminated pet food diets can be colonized by L. monocytogenes without showing clinical signs making the pets a possible source of contamination in the household. This study aimed to detect and enumerate the presence of L. monocytogenes in pet food diets, namely cat and dog food.