Virulence and proteomic responses of Metarhizium anisopliae against Aedes albopictus larvae.

The tropical climate in Malaysia provides an ideal environment for the rapid proliferation of Aedes mosquitoes, notably Aedes aegypti and Aedes albopictus, prominent vectors of dengue fever. Alarmingly, these species are increasingly developing resistance to conventional pesticides. This study aimed to evaluate the efficacy of Metarhizium anisopliae isolate HSAH5 spores, specifically on conidia (CO) and blastospores (BL), against Ae. albopictus larvae. The study centered on evaluating their pathogenic effects and the resultant changes in protein expression. Spore suspensions with varying concentrations were prepared for larvicidal bioassays, and protein expressions were analysed using liquid chromatography-mass spectrometry. Subsequently, protein annotation and network analysis were conducted to elucidate infection mechanisms and the proteomic response. Based on the lethal concentrations and time frames, CO exhibited faster larval mortality than BL at lower concentrations. Despite this, both spore types demonstrated comparable overall pathogenic effects. Results from the proteomic profiling revealed 150 proteins with varied expressions following exposure to Ae. albopictus extract, shedding light on distinct infection strategies between the spores. Gene Ontology enrichment and network analysis illustrated the diverse metabolic adaptations of M. anisopliae and interactions with mosquito larvae. This highlighted the complexity of host-pathogen dynamics and the significance of biosynthetic processes, energy storage, and cellular interaction pathways in disease progression. The BL network, consisting 80 proteins and 74 connections, demonstrates the intricate fungal mechanisms triggered by host stimuli. Conversely, the CO network, though smaller, displayed notable interconnectivity and concentrated involvement at the cell periphery, suggesting a deliberate strategy for initial host contact. This study offers valuable insights into proteome dynamics of M. anisopliae's BL and CO for managing mosquito populations and combating disease transmission, thereby significantly advancing public health and environmental conservation efforts.

Molecular prevalence of Bartonella spp. in bat flies in east coast Malaysia.

Bats are a significant reservoir for numerous pathogens, including Bartonella spp. It is one of the emerging zoonotic bacterial diseases that can be transmitted to humans and may cause various unspecific clinical manifestations. Thus, bartonellosis is rarely diagnosed and is regarded as a neglected vector-borne disease (VBD). Bat flies have been hypothesised to be a vector in the transmission of pathogens among bats. They are host-specific, which reduces the likelihood of pathogen transmission across bat species; however, they are likely to maintain high pathogen loads within their host species. To explore the presence of Bartonella spp. in bat flies from Peninsular Malaysia; bat fly samples collected from various sites at the east coast states were subjected to molecular detection for Bartonella spp. It was discovered that 38.7 % of bats from Terengganu and Kelantan were infested with bat flies; however, no bat fly was found in bats collected from Pahang. The collected bat flies belonged to the families Nycteribiidae (79.6 %) and Streblidae (20.4 %). The collected bat flies were pooled according to the locations and species into 39 pools. Out of these 39 pools, 66.7 % (n = 26) were positive for Bartonella spp. by PCR. Sequence analyses of five randomly selected PCR-positive pools revealed that pools from Kelantan (n = 3) have the closest sequence identities (99 %) to Bartonella spp. strain Lisso-Nig-922 from Nigeria. However, the other pools from Terengganu (n = 2) were closely related to Bartonella spp. strain KP277 from Thailand and Bartonella spp. strain Rhin-3 from the Republic of Georgia with 99 % and 100 % sequence identity, respectively. This suggests that the Bartonella spp. found in Malaysian bat flies are genetically diverse and can potentially serve as reservoirs for pathogenic Bartonella spp.

Survey on tick infestation in stray dogs in localities of Malaysia.

The close relationship between dogs and humans has led to concerns regarding the transmission of zoonotic pathogens through ectoparasites such as ticks. This study aimed to investigate the occurrence of ticks and the intensity on stray dogs with specific risk factors (size, sex, neutering status, body part). Additionally, identifying the predilection sites of ticks on stray dogs was crucial for developing an effective tick control program in Malaysia. A cross-sectional study was conducted among 64 stray dogs from Kelantan and Selangor States. These dogs were subjected to integumentary examinations, collecting 431 ticks comprising Rhipicephalus sanguineus s.l. "tropical lineage" and Haemaphysalis bispinosa from 53 infested dogs. The overall occurrence of tick infestation was 82.81% (53/64), with an average intensity of 8.13 ticks (range: 1-17) per stray dog. All the potential risk factors considered in this study showed no statistically significant result (P value >0.05). The head, ear, and neck were the most preferred attachment sites for ticks. These findings underscore the importance of implementing tick control programs for stray dogs, which serve as reservoirs of ticks and tick-borne pathogens for owned dogs and humans.

The first molecular identification and phylogenetic analysis of tick-borne pathogens in captive wild animals from Lohi Bher zoo, Pakistan.

Tick-borne pathogens are causing severe diseases in livestock, wild animals, and humans. Wild animals play a crucial role in tick-borne pathogens' transmission life cycle by serving as reservoir hosts or intermediate hosts, posing a continuous risk for domestic animals and humans. The presence of tick-borne pathogens is often ignored in wild animals kept in zoos, which is a public health concern. In the present study, we investigated these pathogens in tick-infested captive wild animals at the Lohi Bher zoo, Pakistan. Blood samples were collected from 22 animals, which include urials (4) (Ovis aries vignei), blackbucks (3) (Antilope cervicapra), fallow deer (1) (Dama dama), hog deer (6) (Axis porcinus), chinkaras (4) (Gazella bennettii), white tiger (2) (Panthera tigris tigris), a giraffe (Giraffa camelopardalis), and African lions (2) (Panthera leo). The samples were screened for Piroplasm and Anaplasma spp. by polymerase chain reaction targeting different gene loci. We detected three Theileria spp. and one Anaplasma sp. from the investigated captive wild animals. The Theileria sp. dama gazelle was detected from chinkara, Theileria sp. NG-2012b from chinkara and giraffe and T. parva from African lion, and Anaplasma bovis was identified in a giraffe. Moreover, Theileria sp. and Anaplasma sp. coinfection was detected in one giraffe. Overall, this study shows that Theileria spp. and Anaplasma spp. are circulating in captive wild animals, which can play an important role in their spread. Further studies are required to monitor tick-borne pathogens in zoo animals and their potential to spread from exotic wild captive animals to local wild and domestic.

Antibacterial and cytotoxic activity assessment of Channa striatus (Haruan) extract.

BACKGROUND AND AIM: Channa striatus extract, a freshwater snakehead fish known as Haruan, is popular in Southeast Asia for consumption and as a traditional therapeutic remedy for wound healing. C. striatus is also used in osteoarthritic for its anti-inflammatory. The aim of this study was to determine the presence of antibacterial properties of C. striatus extract against oral bacteria and to investigate the cytotoxic activity against Vero cells. MATERIALS AND METHODS: The authors prepared C. striatus extract in chloroform-methanol solvents. Next, the authors took subgingival microbiological samples from 16 cats that had periodontal disease. The authors determined the antibacterial properties of C. striatus extract against the isolated bacteria using the disk diffusion method and a broth microdilution-based resazurin microtiter assay. Finally, the authors used the Vero cell line to evaluate the cytotoxic activity, and they assessed the cell availability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: The results showed weak antibacterial activity of C. striatus extract against Pseudomonas spp. and Escherichia coli. In addition, the authors found that minimum inhibition concentration values ranged between 400 and 500 mg/mL, and minimum bactericidal concentration values ranged between 650 and 550 mg/mL. However, the cytotoxic results were promising, showing that C. striatus extract increased the cell viability and growth when it was at a higher concentration. The extract also promotes growth and cell proliferation. CONCLUSION: These findings suggest that C. striatus extract promoted cell proliferation in vitro and could be a plausible therapeutic wound healing alternative for periodontal disease in cats.

Prevalence of parasites in working pig-tailed Macaques (Macaca nemestrina) in Kelantan, Malaysia.

BACKGROUND: Coconut is an important commodity in Kelantan, and pig-tailed macaques (Macaca nemestrina) have been traditionally used for coconut-plucking for over a century. Most of these animals were sourced from the wild population, and the parasitic status of these macaques is unknown, plus the impacts caused by these parasites are usually underestimated by the owners. METHODS: A total of 30 macaques were sampled for blood, faeces and hair plucks to detect parasite. RESULTS: Out of 21 faecal samples examined, 11 (52%) were determined positive for one or more gastrointestinal parasites, namely Trichostrongylus spp., Strongyloides spp., Anatrichosoma spp., Capillaria spp., Trichuris spp. and Paramphisotomum spp. Filaria was detected in one (3%) of the blood samples. For ectoparasites, only lice, Pedicinus sp., were found in 9 (30%) macaques. CONCLUSIONS: It is imperative that the parasitic status of these animals be determined so that necessary actions and preventive measures can be implemented to prevent zoonotic transmissions.