Phytochemicals and Nano-Phytopharmaceuticals Use in Skin, Urogenital and Locomotor Disorders: Are We There?

Nanomedicines emerged from nanotechnology and have been introduced to bring advancements in treating multiple diseases. Nano-phytomedicines are synthesized from active phytoconstituents or plant extracts. Advancements in nanotechnology also help in the diagnosis, monitoring, control, and prevention of various diseases. The field of nanomedicine and the improvements of nanoparticles has been of keen interest in multiple industries, including pharmaceutics, diagnostics, electronics, communications, and cosmetics. In herbal medicines, these nanoparticles have several attractive properties that have brought them to the forefront in searching for novel drug delivery systems by enhancing efficacy, bioavailability, and target specificity. The current review investigated various therapeutic applications of different nano-phytopharmaceuticals in locomotor, dermal, reproductive, and urinary tract disorders to enhance bioavailability and efficacy of phytochemicals and herbal extracts in preclinical and in vitro studies. There is a lack of clinical and extensive preclinical studies. The research in this field is expanding but strong evidence on the efficacy of these nano-phytopharmaceuticals for human use is still limited. The long-term efficacy and safety of nano-phytopharmaceuticals must be ensured with priority before these materials emerge as common human therapeutics. Overall, this review provides up-to-date information on related contemporary research on nano-phytopharmaceuticals and nano-extracts in the fields of dermatological, urogenital, and locomotor disorders.

Bacteriological and histopathological findings in cetaceans that stranded in the Philippines from 2017 to 2018.

The relatively high frequency of marine mammal stranding events in the Philippines provide many research opportunities. A select set of stranders (n = 21) from 2017 to 2018 were sampled for bacteriology and histopathology. Pertinent tissues and bacteria were collected from individuals representing eight cetacean species (i.e. Feresa attenuata, Kogia breviceps, Globicephala macrorhynchus, Grampus griseus, Lagenodelphis hosei, Peponocephala electra, Stenella attenuata and Stenella longirostris) and were subjected to histopathological examination and antibiotic resistance screening, respectively. The antibiotic resistance profiles of 24 bacteria (belonging to genera Escherichia, Enterobacter, Klebsiella, Proteus, and Shigella) that were isolated from four cetaceans were determined using 18 antibiotics. All 24 isolates were resistant to at least one antibiotic class, and 79.17% were classified as multiple antibiotic resistant (MAR). The MAR index values of isolates ranged from 0.06 to 0.39 with all the isolates resistant to erythromycin (100%; n = 24) and susceptible to imipenem, doripenem, ciprofloxacin, chloramphenicol, and gentamicin (100%; n = 24). The resistance profiles of these bacteria show the extent of antimicrobial resistance in the marine environment, and may inform medical management decisions during rehabilitation of stranded cetaceans. Due to inadequate gross descriptions and limited data gathered by the responders during the stranding events, the significance of histopathological lesions in association with disease diagnosis in each cetacean stranding or mortality remained inconclusive; however, these histopathological findings may be indicative or contributory to the resulting debility and stress during their strandings. The findings of the study demonstrate the challenges faced by cetacean species in the wild, such as but not limited to, biological pollution through land-sea movement of effluents, fisheries interactions, and anthropogenic activities.

Analysis of dengue cases and severity classifications in Cavite Province, Philippines.

BACKGROUND & OBJECTIVES: Dengue is considered endemic in many countries in the world including the Philippines, and factors associated with dengue infections have not been adequately explored in the Philippines. The aim of this study was to assess demographic and location-related factors associated with different classifications of dengue: mild dengue, dengue fever, and hemorrhage dengue fever. METHODS: This study used consolidated dengue reports of 18482 individual cases from the Provincial Hospital (PH) of Cavite province from 2009-2014 and clinical classifications of dengue used by the Philippine Integrated Disease Surveillance and Response (PIDSR). Multinomial logistic regression and marginal effects were used to analyze factors associated with different dengue classifications. RESULTS: Living in densely populated cities and municipalities, individuals aged 19 years and below, and being female were closely associated with severe dengue (DHF) type, while being male and older (above 19 years old) decreased the risk of contracting severe dengue. INTERPRETATION & CONCLUSION: Our study provides a preliminary assessment of association between demographic factors (gender and age-group), locations (municipalities and cities); and three classifications of dengue (mild, moderate, and severe) in Cavite province in the Philippines. To establish whether dengue is linked to populated areas, age and gender will require further assessments.

Potential anti-Acanthamoeba and anti-adhesion activities of Annona muricata and Combretum trifoliatum extracts and their synergistic effects in combination with chlorhexidine against Acanthamoeba triangularis trophozoites and cysts.

Plants with medicinal properties have been used in the treatment of several infectious diseases, including Acanthamoeba infections. The medicinal properties of Cambodian plant extracts; Annona muricata and Combretum trifoliatum were investigated against Acanthamoeba triangularis. A total of 39 plant extracts were evaluated and, as a result, 22 extracts showed positive anti-Acanthamoeba activity. Of the 22 extracts, 9 and 4 extracts showed anti-Acanthamoeba activity against trophozoites and cysts of A. triangularis, respectively. The minimum inhibitory concentration of A. muricata and C. trifoliatum extracts against trophozoites and cysts was 500 and 1,000 µg/mL, respectively. The combination of A. muricata at 1/4×MIC with chlorhexidine at 1/8×MIC demonstrated a synergistic effect against trophozoites, but partial synergy against cysts. A 40% reduction in trophozoites and 60% of cysts adhered to the plastic surface treated with both extracts at 1/2×MIC were noted comparing to the control (P < 0.05). Furthermore, a reduction of 80% and 90% of trophozoites adhered to the surface was observed after pre-treatment with A. muricata and C. trifoliatum extracts, respectively. A 90% of cysts adhered to the surface was decreased with pre-treatment of A. muricata at 1/2×MIC (P < 0.05). A 75% of trophozoites and cysts from Acanthamoeba adhered to the surface were removed after treatment with both extracts at 4×MIC (P < 0.05). In the model of contact lens, 1 log cells/mL of trophozoites and cysts was significantly decreased post-treatment with both extracts compared to the control. Trophozoites showed strong loss of acanthopodia and thorn-like projection pseudopodia, while cysts demonstrated retraction and folded appearance treated with both extracts when observed by SEM, which suggests the potential benefits of the medicinal plants A. muricata and C. trifoliatum as an option treatment against Acanthamoeba infections.

Acanthamoeba in Southeast Asia - Overview and Challenges.

Acanthamoeba, one of free-living amoebae (FLA), remains a high risk of direct contact with this protozoan parasite which is ubiquitous in nature and man-made environment. This pathogenic FLA can cause sight-threatening amoebic keratitis (AK) and fatal granulomatous amoebic encephalitis (GAE) though these cases may not commonly be reported in our clinical settings. Acanthamoeba has been detected from different environmental sources namely; soil, water, hot-spring, swimming pool, air-conditioner, or contact lens storage cases. The identification of Acanthamoeba is based on morphological appearance and molecular techniques using PCR and DNA sequencing for clinico-epidemiological purposes. Recent treatments have long been ineffective against Acanthamoeba cyst, novel anti-Acanthamoeba agents have therefore been extensively investigated. There are efforts to utilize synthetic chemicals, lead compounds from medicinal plant extracts, and animal products to combat Acanthamoeba infection. Applied nanotechnology, an advanced technology, has shown to enhance the anti-Acanthamoeba activity in the encapsulated nanoparticles leading to new therapeutic options. This review attempts to provide an overview of the available data and studies on the occurrence of pathogenic Acanthamoeba among the Association of Southeast Asian Nations (ASEAN) members with the aim of identifying some potential contributing factors such as distribution, demographic profile of the patients, possible source of the parasite, mode of transmission and treatment. Further, this review attempts to provide future direction for prevention and control of the Acanthamoeba infection.

Impact of Typhoon Haiyan on a Philippine Tarsier Population.

Over the last 2 decades the Philippine tarsier (Carlito syrichta aka Tarsius syrichta) has had its conservation status revised from Endangered to Data Deficient to Near Threatened. The last status change was based on a study of the species' population density, which suggested that a single natural catastrophe could potentially wipe out the Philippine tarsier. In 2013 typhoon Haiyan hit Bohol, one of the island strongholds for this species. In this study we compare the density of the Bohol tarsier population within the Philippine Tarsier and Wildlife Sanctuary before and after the typhoon. We demonstrate that the typhoon significantly affected the density of the Philippine tarsier in the sanctuary. Before the typhoon, tarsier density was approximately 157 individuals/km2 whereas after the typhoon the density was a mere 36 individuals/km2. Prior to the typhoon, more Philippine tarsiers were found in older secondary forest than in younger secondary forest, whereas after the typhoon all observed individuals were found in relatively younger secondary forest. Vegetation plots where we observed Philippine tarsiers prior to the typhoon contained a mean of 33 trees/m2, with a mean diameter at breast height (DBH) of 24 cm, and a mean height of 4 m. After the typhoon vegetation plots contained an average of 156 trees, had a mean DBH of 6 cm, and a mean height of 2 m. Based on the IUCN Red List criteria, the reduction and fluctuation in the density of this species suggests that the conservation status of the Philippine tarsier should be changed to Vulnerable. This study indicates natural disasters can have a significant effect on the extinction risk of primates, with implications for future effects of anthropogenic climate change.

Economic costs and burden of dengue disease in Cavite Province, Philippines / Custos econômicos e impacto da dengue na Província de Cavite, Filipinas

AIMS: To determine the economic costs and the burden of dengue disease in Cavite Province, one of the areas highly inflicted by dengue disease in the Philippines. METHODS: This study used Disability Adjusted Life Years (DALYs) to calculate burden of dengue disease and quantified direct and indirect costs due to hospitalization and ambulatory dengue cases. DALYs were estimated using methods developed by the World Health Organization and the World Bank. We specifically calculated Years of Life Lost from 2009 to 2014 using patient-level data from hospitals and derived Years Lived with Disability from hospital records of dengue specific type (dengue fever/dengue hemorrhagic fever) in Cavite Province. RESULTS: An annual cost of Philippine Peso (PHP) 99,147,173 which is equivalent to United States Dollar (USD) 2,300,000 was obtained. The average annual cost per dengue case was PHP 32,324 (USD 734). The 21-25 age sub-groups had the highest average annual direct cost which amounted to PHP 243,181 (USD 5,526), followed by the 45-54 age sub-groups which amounted to PHP 201,481 (USD 4,579). From 2009 to 2014, the annual burden of disease was estimated at 178,282 DALYs (equivalent to one DALY lost per 17 persons in Cavite Province). CONCLUSIONS: The estimates of costs and DALYs suggested substantial dengue disease burden and economic costs in Cavite Province, Philippines.

OBJETIVOS: Determinar os custos econômicos e o impacto da dengue na Província de Cavite, uma das áreas altamente infligidas por dengue nas Filipinas. MÉTODOS: Este estudo utilizou os anos de vida ajustados por incapacidade (Disability Adjusted Life Years - DALYs) para calcular o impacto da dengue e quantificar os custos diretos e indiretos devidos aos casos de dengue atendidos em hospital ou em ambulatório. Os DALYs foram estimados usando métodos desenvolvidos pela Organização Mundial de Saúde e pelo Banco Mundial. Calculamos especificamente os anos de vida perdidos entre 2009-2014, usando dados dos pacientes hospitalizados. Os anos vividos com incapacidade foram derivados de registros hospitalares sobre o tipo específico da dengue (dengue clássica / dengue hemorrágica) na Província de Cavite. RESULTADOS: Foi identificado um custo anual de 99.147.173 pesos das Filipinas (PHP), equivalentes a 2.300.000 dólares dos Estados Unidos (USD). O custo médio anual por caso de dengue foi 32.324 PHP (734 USD). O subgrupo etário 21-25 anos teve o maior custo direto anual, que atingiu 243.181 PHP (5.526 USD), seguido pelo subgrupo 45-54 anos, que atingiu 201.481 PHP (4.579 USD). De 2009 a 2014, o impacto anual da doença foi estimado em 178,282 DALYs (equivalentes a um DALY pedido por cada 17 pessoas na Província de Cavite). CONCLUSÕES: As estimativas de custos e os DALYs sugerem um alto impacto e substanciais custos econômicos da dengue na Província de Cavite, Filipinas.

Presence of Cryptosporidium parvum and Giardia lamblia in water samples from Southeast Asia: towards an integrated water detection system.

BACKGROUND: Access to clean and safe drinking water that is free from pathogenic protozoan parasites, especially Cryptosporidium parvum and Giardia lamblia that cause gastrointestinal illness in humans, is still an issue in Southeast Asia (SEA). This study is the first attempt to detect the aforementioned protozoan parasites in water samples from countries in SEA, using real-time polymerase chain reaction (qPCR) assays. METHODS: A total of 221 water samples of 10 l each were collected between April and October 2013 from Malaysia (53), Thailand (120), the Philippines (33), and Vietnam (15). A physicochemical analysis was conducted. The water samples were processed in accordance with the US Environmental Protection Agency's methods 1622/1623.1, microscopically observed and subsequently screened using qPCR assays. RESULTS: Cryptosporidium oocysts were detected in treated water samples from the Philippines (1/10), with a concentration of 0.06 ± 0.19 oocyst/L, and untreated water samples from Thailand (25/93), Malaysia (17/44), and the Philippines (11/23), with concentrations ranging from 0.13 ± 0.18 to 0.57 ± 1.41 oocyst/L. Giardia cysts were found in treated water samples from the Philippines (1/10), with a concentration of 0.02 ± 0.06 cyst/L, and in untreated water samples from Thailand (20/93), Vietnam (5/10), Malaysia (22/44), and the Philippines (16/23), with concentrations ranging from 0.12 ± 0.3 to 8.90 ± 19.65 cyst/L. The pathogens C. parvum and G. lamblia were detected using using qPCR assays by targeting the 138-bp fragment and the small subunit gene, respectively. C. parvum was detected in untreated water samples from the Philippines (1/23) and Malaysia (2/44), whilst, G. lamblia detected was detected in treated water samples from the Philippines (1/10) and in untreated water samples from Thailand (21/93), Malaysia (12/44), and the Philippines (17/23). Nitrate concentration was found to have a high positive correlation with (oo)cyst (0.993). CONCLUSION: The presence of (oo)cysts in the water samples means that there is potential risk for zoonotic disease transmission in the studied countries. Detection using qPCR is feasible for quantifying both pathogenic C. parvum and G. lamblia in large water samples.

Knowledge and practice on Toxoplasma infection in pregnant women from Malaysia, Philippines, and Thailand.

Toxoplasma gondii, is one of the infectious agents of congenital TORCH infections, causes severe clinical outcomes in fetus and newborns. Nevertheless this life-threatening parasitic disease is preventable by simple preventive measures related to lifestyle during pregnancy. We aim to study on the knowledge about toxoplasmosis and practices that prevents this infection among the pregnant women. Total of 2598 pregnant women from Malaysia, Philippines, and Thailand were randomly surveyed to determine the knowledge and their practices on Toxoplasma infection. The questionnaire covered respondents' general information and knowledge on plausible risks factors, symptoms, timing of infection, prevention knowledge, and preventive behavior regarding Toxoplasma infection. Majority of these pregnant women were in their age group of 20-29 years (50.9%), completed secondary level of education (51.7%), in their second trimester of pregnancies (38.1%), non-parous (36.6%), and had no history of abortion (90.4%). Based on this survey, only 11% of these pregnant women had read, heard, or seen information regarding toxoplasmosis and 3.5% of them were aware of being tested for the infection. A small percentage of these pregnant women knew that T. gondii were shed in the feces of infected cats (19.4%) and sometimes found in the raw or undercooked meat (11.0%). There was 16.1% of responding women knew that toxoplasmosis is caused by an infection. Demographic profiles such as age group, level of education, pregnancy term, and number of children of the pregnant women showed significant association with their responses toward prevention knowledge and preventive behavior related questions (P < 0.05). Thus, it is suggested that health education on toxoplasmosis and primary behavioral practices should be consistently offered to reproductive age women in general and pregnant women in particular. This information could help to reduce vertical transmission of Toxoplasma infection during pregnancy.

Waterborne parasites: a current status from the Philippines.

BACKGROUND: Despite the amount of awareness created, waterborne disease still poses threat, especially in developing countries. Due to the scarcity of reported data on waterborne parasites, the consumption of unsafe water prolongs. Thus, the occurrences of waterborne parasites from various samples were investigated from one of the Southeast Asian country, the Philippines. METHODS: A total of thirty three samples, each consisting of twelve liters, were collected and processed to obtain the sediment. Ten liters of sample each was processed to detect Cryptosporidium spp. and Giardia spp. using an immunomagnetic separation method prior to enumeration via fluorescence microscope. Meanwhile, the remaining two liters were cultured to detect Acanthamoeba and Naegleria through microscopy examination and polymerase chain reaction (PCR) analysis. RESULTS: Twelve samples (36.4%) from river (5), swimming pool (1), pond (3), rain tank (1), and natural lake (2) were positive for Cryptosporidium spp., 17 (45.5%) samples from river (9), pond (2), swimming pool (1), rain tank (1), and natural lake (4) were positive for Giardia spp. while, 13 (33.3%) samples from river (3), swimming pool (2), pond (2), dispenser (1), well (1), tap (2) and natural lake (2) were positive for Acanthamoeba spp. and 5 (18.2%) samples from river (1), natural lake (1), tap (1), dispenser (1) and mineral (1) were Naegleria spp. positive. Physical parameters such as temperature, conductivity, total dissolved solid (TDS), salinity, dissolved oxygen (DO), pH, and turbidity and chemical parameters such as ammonia, chlorine, fluoride, nitrate and nitrite were also measured. The highest chemical contamination was observed at pond 2. A good correlation was observed between Giardia and nitrite (r = 0.736, p < 0.01) and Giardia and nitrate (r = 0.502, p < 0.01). CONCLUSION: This study was aimed to create greater awareness of parasitic contamination in the water environment in the Philippines and also to act as a platform of the current scenario for policymakers as water pollution is a key health issue in this region.

Comparative study on waterborne parasites between Malaysia and Thailand: A new insight.

This study investigated the distribution of parasites as main contaminants in water environments of peninsular Malaysia (October 2011-December 2011) and the southeastern coast of Thailand (June 2012). Sixty-four water samples, 33 from Malaysia and 31 from Thailand, of various water types were examined according to U.S. Environmental Protection Agency guidelines. Drinking or household water types from both countries were free from parasitic contamination. The recreational/environmental (except a swimming pool in Malaysia) and effluent water types from these two countries were contaminated with waterborne parasites: Giardia (0.04-4 cysts/L), Cryptosporidium (0.06-2.33 oocysts/L), hookworm (6.67-350 ova/L), Ascaris (0.33-33.33 ova/L), and Schistosoma (9.25-13.33 ova/L). The most contaminated sites were recreational lake garden 3 in Malaysia and river 2 in Thailand. Higher concentrations of Giardia, Cryptosporidium, and hookworm were found in samples from Malaysia than in samples from Thailand. The presence of Giardia cysts showed a significant association with the presence of Cryptosporidium oocysts (P < 0.005).

Waterborne parasites and physico-chemical assessment of selected lakes in Malaysia.

The objective of this study was to assess the physico-chemical parameters and waterborne parasites in selected recreational lakes from Malaysia. Samples were collected from seven stations of Recreational Lake A (RL-A) and six stations of Recreational Lake B (RL-B). The samples were processed to detect the presence of Giardia spp. and Cryptosporidium spp. using immunomagnetic separation kit, helminth eggs or ova by bright field microscopy and Acanthamoeba spp. by cultivation in non-nutrient agar. Chemical parameters such as ammonia, chlorine, fluoride, nitrate and nitrite and physical parameters such as dissolved oxygen, electrical conductivity, pH, salinity, temperature and total dissolved solid were also measured. Both lakes were freshwater with salinity ranging from 0.05 to 0.09 ppt. Most stations of these lakes were contaminated with Cryptosporidium spp., Giardia spp., Ascaris spp. and hookworm. Schistosoma spp. was found in RL-B only, while Acanthamoeba spp. was found in all stations. Of all sampling sites, station 5 of RL-B is the most contaminated. Linear regression and correlation analysis revealed that Giardia spp. and Schistosoma spp. showed a significant negative correlation with turbidity (p < 0.01). Based on the preliminary data obtained, it is clearly shown that there is a necessity to implement the detection of waterborne parasites and physico-chemical analysis in Malaysia. Future work on heavy metals (chromium, copper, mercury and zinc) is recommended to enhance the overall water quality monitoring and to take appropriate safety measures to ensure maintenance of good water standards.

Detecção sorológica da infecção por Toxoplasma gondii em gatos errantes e domiciliados e sua avaliação hematológica / Serologic detection of toxoplasma gondii infection in stray and household cats and its hematologic evaluation

Aims: This study focused on the serologic detection of Toxoplasma gondii infection in two groups of cats:stray and household groups. In addition, hematologic assessment of seropositive and seronegative cats was done. Methods: Sixty cats were serologically tested for anti-Toxoplasma gondii antibodies using the latex agglutination test. Six collection sites for each group of cats were identified in the urban communities of Sta Rosa and San Pedro, Laguna, Philippines. The 60 cats collected were divided into 30 stray and 30 household cats. Results: Results revealed that 28 (46.67%) of the 60 cats were seropositive. There were more household cats (28.33%) which showed seropositivity compared to stray cats (18.33%), however the difference was statistically insignificant (p>0.05) . Hematologic tests through complete blood count showed significantly (p<0.05) higher number of seropositive cats with abnormalities on hemoglobin level, red blood cell count, segmenter (neutrophil) and monocyte counts compared to the control. Other parameters such as percent packed cell volume, white blood cell count, eosinophil and lymphocyte counts showed insignificant (p>0.05) results across seropositive cats and the control. Blood chemistry analysis showed significantly higher (p<0.05) potassium level irregularities in seropositive cats relative to the seronegative cats. Other parameters such as amylase, blood sugar, blood uric acid, creatinine and blood urea nitrogen were statistically insignificant (p>0.05). Conclusions: Although Toxoplasma gondii infection suggests possible cause of hematologic abnormalities, it is recommended that further studies on this aspect be done to provide more basic and clinical research information that would improve cat health management.

Serologic detection of Toxoplasma gondii infection in Rattus spp collected from three different sites in Dasmariñas, Cavite, Philippines.

Acute and chronic cases of toxoplasmosis in Rattus norvegicus and Rattus rattus mindanensis caught in agricultural, commercial and residential sites in Dasmariñas, Cavite, Philippines were determined serologically. Fifty-eight percent of R. norvegicus and 42.0% of R. r. mindanensis were positive for anti-T. gondii antibodies (Abs). Infection was higher in male rats, and those caught in the commercial site had 100.0% seropositivity. Thirty percent of the R. norvegicus and 51.0% R. rattus mindanensis had acute infection, with 1:64-1:128 Abs titer. Seventy percent of the R. norvegicus and 49.0% of R. rattus mindanensis were chronically-infected with Abs titer 1:256-1:2048 and 1:256-1024, respectively. The association between the presence of infection with the rat gender and species and their collection sites was insignificant (p>0.05). In a related study, however, mice experimentally-inoculated brain tissue homogenate obtained from chronically-infected Rattus spp, manifested differences in the onset as well as, severity of infection which was histopathologically evaluated, suggestive of a possible difference in T. gondii parasite strain(s) infecting different rat populations.

Parasite biodiversity in Rattus spp caught in wet markets.

Rattus spp trapped in wet markets in Quiapo, Manila and Balayan, Batangas had ectoparasites, Echinolaelaps echidnius (mite), and Polyplax spinulosa (louse). The endoparasites identified were Hymenolepis diminuta; the acanthocephalan Moniliformis moniliformis; Taenia taeniaeformis strobilocercus larvae and Capillaria hepatica in liver; Trichosomoides crassicauda of the urinary bladder; Sarcocystis sp of muscle tissue; and two different species of stronglyloid-looking intestinal nematodes. Rats had 100% infection with C. hepatica and T. taeniaeformis, exhibiting high parasitemia. The co-existence of rats with diverse parasitic species is reflective of the host's capability to support parasites' behavioral, physiological, and developmental needs. Despite heavy infection with intestinal parasites, and marked hepatic tissue damage owing to severe capillariasis and strobilocercus larval infection, all rats appeared healthy and agile, suggestive of a well-established rat host-parasite relationship. In view of the diversity and zoonotic nature of rat parasites, and the impoverished conditions prevailing in communities where Rattus spp survive and proliferate, they can readily facilitate parasite transmission to humans and other susceptible animal hosts.