Detection of Trypanosoma evansi in a naturally infected cat in Indonesia using bioassay and molecular techniques.

Background and Aim: The prevalence of surra in domestic cat is seldom and it is caused by Trypanosoma brucei and Trypanosoma evansi. However, molecular diagnostic approaches are required owing to similarities in their morphology. In Yogyakarta, a domestic cat was diagnosed with trypanosomiasis; however, the causative species was undetermined. Therefore, we aimed to molecularly and biologically identify the isolate. Materials and Methods: Approximately 1 mL of blood from an infected cat was collected into EDTA tube and separated for inoculation into donor mice, blood smear, and DNA isolation. Two donor mice was then used for increasing the number of parasite in order to infect 10 experimental mice. Parasitemia was monitored daily in each experimental mouse by preparing a wet mount and Giemsa-stained thin blood smear. The blood of experimental mice that reached the peak of parasitemia was then collected and used for DNA isolation. Each blood sample, which collected from infected cat and experimental mice, was then isolated and amplified the DNA by polymerase chain reaction using ITS-1. The parasitemia pattern and viability of the animals were observed to determine the biological characteristics of trypanosomatid, while to assess the molecular characteristics, the internal transcribed spacer (ITS)-1 amplification was used. Results: The prepatent period of this trypanosomatid is between 2 and 4 dpi, whereas the life span of mice is approximately 4-10 dpi. Morphologically, the trypomastigote in the cat blood smear had long slender and intermediate shapes. However, only the long slender form was detected. Among the total of 410 nucleotides (NT) of ITS-1 sequences, 25 NT substitutions differed between the cat and mouse isolates. Phylogenetic analysis revealed that both samples had a close genetic relationship with T. evansi. Conclusion: Trypanosoma evansi, a highly virulent trypanosomatid, was isolated from a cat in Yogyakarta.

Molecular and morphological identification of Lernaea spp. in Cyprinid fishes from two districts in Yogyakarta, Indonesia.

Background and Aim: Parasitic infection commonly affects freshwater ornamental fishes. Parasites in fish may impede their growth and even cause death, resulting in a decline in fecundity. The prevalence of lernaeosis in aquaculture ponds in Indonesia requires attention because of missing data, especially from Yogyakarta. Therefore, this study aimed to identify the Lernaea species found in fish in Indonesia, particularly in Yogyakarta, molecularly and morphologically, as well as an overview of their distribution and the water condition they inhabit. Materials and Methods: Lernaea species were collected from three different fish species in two districts of Yogyakarta, Indonesia, for precise identification. Lernaea specimens were characterized morphologically and subjected to molecular identification based on 18S rRNA and 28S rRNA genes. Results: Lernaea in this study was morphologically and genetically confirmed as Lernaea cyprinacea, and the infection rate in each fish species was different. Water conditions might have contributed to the differences in infection levels. Conclusion: This study characterized L. cyprinacea isolated from Yogyakarta. Future research should focus on sequencing as much molecular information as possible and carrying out more experimental infections.

Development of a multiplex polymerase chain reaction technique for detection and discrimination of Eimeria spp. in cattle in Indonesia.

Background and Aim: Bovine eimeriosis is a disease caused by apicomplexan parasites of the genus Eimeria. It is one of the most important and widespread bovine illnesses in the world. Some of the identified species of bovine eimeriosis have morphologically similar oocysts that are difficult to differentiate. For the identification of particular Eimeria spp., diagnostic laboratories are increasingly turning to DNA-based technology. This study aims to develop a multiplex polymerase chain reaction (mPCR) technique based on the internal transcribed spacer-1 (ITS-1) gene for the simultaneous identification of pathogenic Eimeria spp. in cattle from Sulawesi Island, Indonesia. Materials and Methods: Genomic DNA was extracted by the DNAzol reagent from the purified Eimeria oocysts. Species-specific primers targeting the ITS-1 region were used to amplify the distinct Eimeria spp. Results: Using PCR ITS-1, this study showed that 36 of 120 fecal samples (30%) were infected by Eimeria spp. The multiplex PCR assay allowed for the simultaneous identification of six major Eimeria spp. in a single-tube reaction. The proportion of mixed Eimeria spp. infections was 100% (36/36). The maximum number of Eimeria spp. was five, and the minimum number was two. Conclusion: Identification of six pathogenic Eimeria spp. in cattle was successfully carried out by nested multiplex PCR using ITS-1 gene. In the future, a procedure to detect pathogenic Eimeria spp. in one tube reaction will offer economical and save diagnostic time.

Molecular identification of cercaria Fasciola gigantica in lymnaeid snails in Kulon Progo, Yogyakarta.

Lymnaeid snails play an essential role in transmitting fasciolosis as intermediate hosts. Therefore, this study aims to use the molecular method to identify liver fluke in lymnaeid snails. A total of 320 lymnaeid snails were collected from a rice field. The samples were dissected to collect cercaria and identified using polymerase chain reaction. Furthermore, the internal transcribed spacer 2 (ITS2) was used as the target gene to identify the species of cercaria. The result showed that 3.75% (12/320) of the snails were infected by Fasciola gigantica, while the phylogenetic tree based on ITS2 showed that the cercaria in this study was monophyletic and similar to species from several countries in Southeast Asia, including China. Furthermore, the haplotype network showed that all four cercaria samples were similar with sequences from several countries. This study suggests that the F. gigantica cercaria isolated from lymnaeid snails in Kulon Progo, Yogyakarta, Indonesia, has a sequence similar to that of other species in Southeast Asian countries, although no hybrid type was detected in these sequences. This is the first report on the molecular identification of cercaria F. gigantica isolated from lymnaeid snails in Yogyakarta, Indonesia.

Prevalence and risk factors associated with Eimeria species infection in cattle of different geographical regions of Indonesia.

BACKGROUND AND AIM: Eimeria spp. are gastrointestinal protozoans that affect animal productivity, thereby causing symptoms that range from bloody diarrhea to death. These symptoms cause economic losses to farmers. The distribution of Eimeria spp. in cattle has, therefore, been reported to have spread widely, especially in the tropics and subtropics. Indonesia is a tropical country at high risk of Eimeria infections. This study aimed to identify the prevalence and risk factors related to the levels of eimeriosis in beef cattle originating from different geographic areas in Indonesia. MATERIALS AND METHODS: Here, 817 fecal samples were collected from beef cattle in Indonesia, including 282 calves, 535 adults, 530 males, and 287 females. In addition, 156 semi-intensively and 661 intensively managed cattle were randomly collected. Then, fecal samples were analyzed by parasitology examinations. RESULTS: Screening examination using the sugar flotation modification method showed that Eimeria spp. were prevalent in Indonesia, as 65.4% of the bacterial strain was detected. The prevalence of identified Eimeria spp. in Indonesia was highest in North Maluku (Maluku Island) (94.1%), whereas the lowest levels were observed in West Java (24.0%) (Java Island). The prevalence was also found to be higher in males (79.3%) than females (51.9%). Similarly, levels in semi-intensively managed cattle (66.7%) were higher than those subjected to intensive management (65.9%). However, its prevalence in calf and adult cattle was similar. CONCLUSION: Bovine eimeriosis spp. were detected at high prevalence in Indonesia, and high-level risks were observed in infected males, including those under the semi-intensive management. In addition, although the results from oocyst examinations were based on qualitative analysis, the endemicity levels of Eimeria spp. among farms in Indonesia should be considered because Eimeria spp. were distributed in most parts of Indonesia. Based on the results of this study, we provide the first information about the prevalence of bovine eimeriosis from different geographical locations in Indonesia, which have differing climates associated with the level of the existing risk factors. Hence, farmers are advised to pay more attention to strict biosecurity techniques on their farms, thereby favoring the early control of bovine eimeriosis.

Anthelmintic effect of Indigofera tinctoria L on Haemonchus contortus obtained from sheep in Indonesia.

BACKGROUND AND AIM: Hemonchosis has resulted in huge economic losses for sheep farmers worldwide. Secondary metabolite compounds from Indigofera tinctoria L. can be used as anthelmintics. This study aimed to evaluate the in vitro and in vivo effects of I. tinctoria L. aqueous extract (IAE) as an anthelmintic against adult Haemonchus contortus isolated from sheep. MATERIALS AND METHODS: Ten active adult worms were placed in each Petri dish containing 25 mL of IAE, each having a different concentration of IAE (at concentrations 100, 120, 140, 160, 180, 200, and 220 mg/mL). Each experiment was repeated. The positive control used 1% albendazole, and the negative control used 0.62% saline water. The number of immobile worms and the time of mortality were recorded after 2, 4, 6, and 8 h. The dead worms were subsequently tested using scanning electron microscopy (SEM) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In the in vivo study, 15 sheep with an average fecal egg count (FEC) of 1000 eggs per gram were treated with IAE for 9 days: Group A (negative control, saline water 0.62%), Group B (21 mg/mL), Group C (41 mg/mL), Group D (62 mg/mL), and Group E (positive control, albendazole 100 mg/mL). Measurements of the body weight, FEC reduction test, and hematology testing were performed on days 0, 7, and 14. SEM was performed using worms found from the abomasum of slaughtered sheep. RESULTS: The leaves of I. tinctoria L. contained a number of secondary metabolites, including total tannins, saponins, flavonoids, and alkaloids. The most effective concentration that killed the adult H. contortus worms was 220 mg/mL (93.33% mortality) after 8 h of treatment. The electrophoresis results showed that the protein band at a dose of 22% was less than that of the control. The highest FECR value of the treatment group on the 14th day after treatment was at a dose of 62 mg/mL. The highest weight gain as well as the highest increased hemoglobin (Hb), packed cell volume (PCV), and total erythrocyte count (TEC) values on the 14th day after treatment were at a dose of 41 mg/mL. The SEM results showed that IAE treatment caused the worms' anterior parts to become wrinkled with thick creases and cuticle abrasion (in vitro) and the anterior part to shrink along with the presence of aggregates in the worm cuticle (in vivo). CONCLUSION: The aqueous extract of I. tinctoria contains tannins, saponins, flavonoids, and alkaloids and has an anthelmintic effect with decreased FEC, increased weight gain, Hb, PCV, and TEC, causing damage to the worms' body and reducing the protein profile of adult H. contortus worms.

In vitro and in vivo Areca catechu crude aqueous extract as an anthelmintic against Ascaridia galli infection in chickens.

AIM: Intestinal parasites cause severe diseases at poultry farms, especially in developing countries, including Indonesia. Infections cause significant decreases in productivity at poultry farms. Ascaridia galli is a common nematode found in chickens with a prevalence of up to 92%. This study evaluates the in vitro and in vivo effects of Areca catechu crude aqueous extract (AAE) as an anthelmintic against A. galli. MATERIALS AND METHODS: In the in vitro study, already-infected slaughtered chickens were collected from local slaughterhouses in Yogyakarta. The chicken intestines were carefully examined and transported to the Parasitological Laboratory of Gadjah Mada University, Yogyakarta. A. galli was collected from the intestines and put into a Petri dish containing 0.62% saline solution. Sixty A. galli individuals were soaked in different concentrations of the AAE for 3-6 h. The number of deceased A. galli was recorded by ascertaining whether they had ceased movement using a stereo microscope. The morphological characteristics of the parasites were studied. The in vivo study used 40 female chickens that were 1.5 months old and infected with the eggs of A. galli. Approximately 3000 infective eggs were found in each chicken. There were four treatment groups; Group A served as the control and was treated with Aquadestilata, Group B was treated with 26 mg/ml of AAE, Group C was treated with 79 mg/ml of AAE, and Group D was treated with 50 mg/ml of pyrantel pamoate. The AAE was applied using a pipette for 14 days. The observed parameters include the effect of the treatments on hematology, body weight (BW), and the number of A. galli eggs within each gram of fecal matter. RESULTS: The AAE damaged the morphology, including the anterior end, posterior end, and vulva, of A. galli in vitro. The tegument of the anterior and posterior ends was slightly rougher, wrinkled, and damaged. The vulva and spicule ducts were irregular and indistinct. The average eggs per gram (EPG) in the untreated control increased in vivo from 1450±355.90 to 1975±325.96. The average EPG decreased from 1485±386.62 to 0±0.00 during 14 days of treatment of 79 mg/ml of AAE. The mean BW gain in the treated chickens was significantly (p<0.5) higher than the control. The treatment with 79 mg/ml of AAE increased the total erythrocyte count, total leukocyte count, hemoglobin, and packed cell volume values in the chickens. CONCLUSION: AAE can cause morphological changes and subsequent death in adult A. galli in vitro and effectively decrease ascariasis severity in chickens, therefore increasing overall BW in vivo.

The prevalence of horse trypanosomiasis in Sumba Island, Indonesia and its detection using card agglutination tests.

BACKGROUND AND AIM: Horses have a strategic and vital role to play in the lives of the people of Sumba Island, East Nusa Tenggara Province. They act as social animals that are involved in death ceremonies, horse races, and during pasola, thereby supporting tourism, and are given away as dowry in wedding ceremonies. This study aimed to investigate the prevalence of trypanosomiasis among horses in four districts of Sumba Island by examining clinical symptoms and detecting parasites, antibodies, and other factors that are related to Trypanosoma evansi infection in horses. MATERIALS AND METHODS: We studied a total of 211 horses that belonged to 88 clinical hobby breeders. Giemsa-colored smears and serum were examined in order to detect antibodies using card-agglutination tests (CATT). The study was conducted during the rainy season that lasted from January to March 2017. Potential risk factors such as the species, sex, origin of the livestock, how the livestock were maintained, and the farmers' knowledge concerning trypanosomiasis were recorded using questionnaires. Data were collected annually for three years from 2010-2012 and repeatedly analyzed by a Chi-square test. RESULTS: Clinical signs of trypanosomiasis were found in 34 horses; blood smears were examined using Giemsa staining and negative preparations were obtained at a frequency of 0.0% (0/211). The CATT results generally showed that 13.3% (28/211) of the samples were seropositive for antibodies to T. evansi; the highest percentage, 16.67% (8/48), of seropositivity was found in the West Sumba District, and the lowest, 12.0% (5/50), was found in Southwest Sumba. The incidence of trypanosomiasis was higher (75% [21/28]) among female hip horses; horses with 1-5 years of experience were more susceptible to a T. evansi infection (46.4% [13/28]). In general, farmers on Sumba Island knew of trypanosomiasis (89.8% [79/88]), and 69.3% (61/88) of the farmers reported that their livestock was sick. This study was the first serological study conducted on trypanosomiasis in horses of Sumba Island after the surra outbreak in 2010-2012. There were 3% of farmers who were willing to provide the government with information on implementing a prevention program and controlling the spread of surra on the island. CONCLUSION: The diagnoses of surra disease were made based on clinical symptoms and parasitological examinations. CATTs could be used to diagnose T. evansi infection in horses.

Trematodiasis occurrence in cattle along the Progo River, Yogyakarta, Indonesia.

AIM: This study aimed to measure the occurrence of trematodiasis in cattle along the Progo River, a district of Yogyakarta, Indonesia. The findings help to establish the magnitude of the disease and encourage prevention and treatment of this condition. MATERIALS AND METHODS: Trematode eggs were extracted from 100 fecal samples collected from cattle. The eggs were examined using the sedimentation technique, and the method of Parfitt and Banks was used to differentiate Paramphistomum spp. eggs from Fasciola spp. eggs. RESULTS: The infection rate of trematode parasites was 50%. Cattle experienced multiple infections of both Paramphistomum spp. and Fasciola spp., as well as single infections of one species or the other. All breeds were vulnerable to infections of both trematode species, although different cattle breeds, including Peranakan Ongole crossbreeds, Simmental crossbreeds, and Limousin crossbreeds, showed differences in infection rate. The highest rate of infection with Paramphistomum spp. (15.78%) occurred in the Simmental crossbreeds. The highest rate of infection (31.57%) with Fasciola spp. was in the Peranakan Ongole crossbreeds. Multiple infections of both Paramphistomum spp. and Fasciola spp. were highest in Simmental crossbreed cattle (28.97%). CONCLUSION: The high infection rates of trematode parasites found in fecal samples, particularly of Fasciola spp., indicate that the cattle along the Progo River in Indonesia experience a high rate of trematodiasis disease.

Ticks (Acari: Ixodidae) infestation on cattle in various regions in Indonesia.

BACKGROUND AND AIM: Ticks (Ixodidae) not only cause blood loss in cattle but also serve as vectors for various diseases, thus causing direct and indirect losses. Moreover, tick infestation can cause significant economic losses. This study aimed to identify the diverse species of ticks infesting cattle in five different regions in Indonesia. MATERIALS AND METHODS: Tick specimens were obtained from local cattle in five different areas in Indonesia. The morphology of the specimens was macroscopically and microscopically evaluated, and the resulting data were descriptively and qualitatively analyzed. RESULTS: In total, 1575 ticks were successfully collected from 26 animals. In total, two genera and three species, namely, Rhipicephalus microplus, Haemaphysalis bispinosa, and Rhipicephalus pilans, were identified. The cattle in Yogyakarta and Riau were infested by H. bispinosa, while the cattle in Sukabumi, Bali, and Lombok were infested by R. microplus and R. pilans. The level of infestation varied among regions, with R. microplus being the most commonly found species. CONCLUSION: The results of this study revealed that cattle in different regions of Indonesia were infested by variable numbers of tick species. In particular, the cattle in Yogyakarta and Riau were solely infested by H. bispinosa; this is a new finding in terms of the distribution of tick species in the country. Increased tick infestation in cattle decreases productivity and causes health problems; therefore, it deserves serious attention. Our findings can help in the formulation of an effective strategy for controlling and preventing cattle tick infestation in the country.

Lice infestation and diversity in turkeys (Meleagris gallopavo) in the Special Region of Yogyakarta and Central Java, Indonesia.

BACKGROUND AND AIM: Biting lice (Phthiraptera: Amblycera and Ischnocera) are ectoparasites that play important roles in the transmission of disease agents that infect turkeys and impact turkey productivity. This study aimed to determine the diversity of lice that infest turkeys in the Central Java Province and the Special Region of Yogyakarta, Indonesia. MATERIALS AND METHODS: Lice sampling was conducted at 16 different locations from April 2019 to June 2019 in turkeys aged 4 months to 2 years. The samples were stored in 70% alcohol and were identified using avian louse keys. The morphology of the specimens was macroscopically and microscopically evaluated, and the resulting data were descriptively and qualitatively analyzed. RESULTS: A total of 2505 lice were collected, and two families and five genera of lice were identified. Three lice genus members of the Philopteridae family (Lipeurus, Oxylipeurus, and Chelopistes) and two genera of the Menoponidae family (Colpocephalum and Menacanthus) were identified. Lipeurus was the most frequently identified genera in turkeys, whereas Menacanthus was the most rarely identified one. The White Holland breed had the highest number of lice infestations, whereas the Jersey Buff breed exhibited the highest diversity of lice genera. The average number of lice infestations was higher in male turkeys than in female turkeys. CONCLUSION: The occurrence of ectoparasites in domestic turkeys indicates that the existence and diversity of lice genera in the study location can be influenced by turkey type, turkey maintenance system, enclosure sanitation measures, lack of strategic ectoparasite control, and environmental factors.

Detection of Toxoplasma gondii copro-prevalence by polymerase chain reaction using repetitive 529 bp gene in feces of pet cats (Felis catus) in Yogyakarta, Indonesia.

AIM: The aim of this research was to determine the copro-prevalence of Toxoplasma gondii using polymerase chain reaction (PCR) with repetitive 529 bp gene and to construct the phylogenetic tree of Toxoplasma oocyst from pet cats in Yogyakarta. MATERIALS AND METHODS: 9 of 132 pet cat samples which serologically positive for Toxoplasma were used in this research. To determine the copro-prevalence of T. gondii in pet cat, 10 g of feces samples taken from practitioners and household cats in Yogyakarta were used in the PCR method utilizing repetitive 529 bp gene sequences. RESULTS: The result shows that copro-prevalence by PCR using repetitive 529 bp gene was 33.3% (3/9). The phylogenetic tree of Toxoplasma grouped into two clades, which clade 1 consists of Toxoplasma isolates collected from pet cats in Yogyakarta Indonesia and T. gondii isolates from China and in clade 2 consist of the T. gondii isolates from India. CONCLUSION: Copro-prevalence of T. gondii in pet cats in Yogyakarta by means of PCR using repetitive 529 bp gene is around 33.3%.

Morphology and morphometry of Haemonchus contortus exposed to Gigantochloa apus crude aqueous extract.

AIM: Haemonchus contortus is the most pathogenic nematode infesting the digestive tract of goats and sheep worldwide leading to a tremendous loss in a variety of routes. Economic losses due to haemonchosis in subtropic and tropic areas are usually caused by poor weight gain, minimized growth, loss of production, and mortality. The prevalence of haemonchosis in Indonesia is 89.4% in goat, and annual loss achieved 1 million US dollars. This study evaluated in vitro effects of Gigantochloa apus crude aqueous extract as an anthelmintic on H. contortus morphology and morphometry. MATERIALS AND METHODS: Bligon goats which are naturally infected were collected from slaughtered goat from local slaughterhouses, namely Besi Sleman. Bligon goat's abomasum part was carefully examined and transported to the Parasitology Laboratory, University of Gadjah Mada, Yogyakarta. H. contortus was obtained from 4 to 6-month-old female goat from slaughterhouses in Yogyakarta area. H. contortus was collected from abomasum and put into a Petri dish containing 0.62% water saline. The number of H. contortus used for each concentration is 25. H. contortus was soaked in each concentration for 4 h. The figure of the parasites or parts of parasites was captured using camera Lucida, and they were measured using both objective micrometer and objective ocular micrometer. All the capturing processes were done with the help of Olympus Digital Camera under Olympus CX21 microscopic. Parasite morphology was identified in morphological and morphometric characters. RESULT: Morphology of H. contortus revealed the cervical papillae bulge appears unclear shape and anterior end is more tapered. Vulvar flab control is not tapered, but vulvar flab which gets aware of G. apus crude aqueous extract looks more pointed. The gubernaculum appears irregular compared to gubernaculum control which tends to be more compact, and the posterior end form appears irregular more than posterior end control. Morphometry study of H. contortus indicates that it has a significant difference for body length, body width, cervical papillae, and spicule length in the male. CONCLUSION: G. apus crude aqueous extract activity revealed morphology change such as cervical papillae, vulvar flab, gubernaculum, posterior end, and reduced morphometry measurement of H. contortus adult worms, notably in body length, body width, cervical papillae width, gubernaculum, and spicule length in males and body length, body width, cervical papillae width, and vulva length in females.

In vitro potential anthelmintic activity of Biophytum petersianum on Haemonchus contortus.

AIM: Haemonchus contortus is a major problem in small ruminants in Indonesia. The frequent use of the anthelmintic drugs has given rise to drug-resistant populations which increase the need for new anthelminthic compounds, particularly from endemic plants. This study evaluated the in vitro effects of Biophytum petersianum crude aqueous extract (BAE) as an anthelmintic compound against H. contortus adult worm isolated from goats. MATERIALS AND METHODS: Adult worm collected from naturally infected abomasums were obtained from slaughtered goats on the day of slaughter. BAE was prepared in six different concentrations (10, 20, 40, 60, 80, and 100 mg/ml) which were tested for their efficacies on ten actively moving worms. Ivermectin (1 mg/ml) was included as a reference drug, while saline water was included as a control. The dead worms from anthelmintic test then went through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and scanning electron microscopy (SEM). RESULTS: Highest mean mortality in treatments group both at 2 h and 4 h observations was BAE 10%. The SDS-PAGE analysis revealed the presence of five protein bands with molecular weights 9.3, 17.1, 50.0, 63.2, and 72.7 kDa based on BAE 10%. The SEM changes observed in the in vitro trials revealed the occurrence of interactions between the BAE and the cuticle. CONCLUSION: The SEM and SDS-PAGE analysis revealed ultrastructural structural changes and the decrease numbers of polypeptides on treated worms when compared to the control worms. It can thus be concluded that the BAE exhibits good anthelmintic activity against H. contortus adult worm.

Aedes aegypti resistance development to commonly used insecticides in Jakarta, Indonesia.

Aedes aegypti is the primary vector of various relevant arthropod-borne viral infectious diseases worldwide. The mosquito control is still mainly performed by using insecticides but their effectiveness is increasingly questioned nowadays. We here conducted a study on Ae. aegypti resistance development towards several commonly used insecticides in the capital city of Jakarta, Indonesia. In order to achieve this goal, Ae. aegypti eggs from Jakarta were collected with ovitraps and hatched in the insectary of the Gadjah Mada University, Indonesia. The F0 generations were used for WHO resistance tests and knockdown resistance (kdr) assays. Presented results clearly showed that there was resistance development of Ae. aegypti populations to the here tested pyrethroid insecticides (i. e. permethrin). Observed mortalities were less than 90% with highest resistance against 0.75% permethrin concentrations. Furthermore, a significant association of V1016G gene mutations with resistance phenotypes to 0.75% permethrin was observed. Nevertheless, the F1534C mutation did not show a significant correlation to resistance development. In conclusion, our results show that populations of Ae. aegypti mosquitoes within the city of Jakarta have developed resistance against several routinely used pyrethroid insecticides in local performed control programs. Thus, the regular verification/assessment of resistance development status will hopefully help in the future to assist local public health authorities in their mosquito control programs by recommending and managing the rotation of different routinely used insecticides with diverse effector mechanisms in order to delay Ae. aegypti resistance development.

Detection of Toxoplasma gondii in cat's internal organs by immunohistochemistry methods labeled with-[strept] avidin-biotin.

AIM: The aims of the study are to detect the presence of Toxoplasma gondii antigen and to determine its distribution location in several organs of domestic cat using immunohistochemistry (IHC) method with Labeled-[Strept] Avidin-Biotin (LAB-SA). MATERIAL AND METHODS: Four domestic cats aged 1-2 years were used as sample in this research. The sample divided into two groups with two cats each. Cats in Group I were positive Toxoplasma based on serologically screening test, while cats in Group II were orally infected with 1×106Toxoplasmaoocyst. All samples then necropsied, and the organs including brain, liver, kidney, duodenum, jejunum, ileum, lungs, and spleen were collected for IHC method with LAB-SA. RESULT: The result showed that Toxoplasma antigens were detected in ileum of both serologically positive domestic cat and the experimentally infected cats. Toxoplasma was also observed in kidney of serologically positive domestic cat. In the serologically positive domestic cat, necrotic lesions were found on ileum, kidney, and liver, whereas in experimentally infected cat, the lesion was only found on ileum. CONCLUSION: The presence of Toxoplasma antigen is successfully detected in several organs of domestic cat using IHC method with the LAB-SA.

Knockdown resistance (kdr) of the voltage-gated sodium channel gene of Aedes aegypti population in Denpasar, Bali, Indonesia.

BACKGROUND: Aedes aegypti is the main vector of several arthropod-borne viral infections in the tropics profoundly affecting humans, such as dengue fever (DF), West Nile (WN), chikungunya and more recently Zika. Eradication of Aedes still largely depends on insecticides, which is the most cost-effective strategy, and often inefficient due to resistance development in exposed Aedes populations. We here conducted a study of Ae. aegypti resistance towards several insecticides regularly used in the city of Denpasar, Bali, Indonesia. METHODS: Aedes aegypti egg samples were collected with ovitraps and thereafter hatched in the insectary of the Gadjah Mada University. The F0 generation was used for all bioassay-related experiments and knockdown resistance (kdr) assays. RESULTS: Results clearly showed resistance development of Ae. aegypti against tested insecticides. Mortalities of Ae. aegypti were less than 90% with highest resistance observed against 0.75% permethrin. Mosquitoes from the southern parts of Denpasar presented high level of resistance pattern in comparison to those from the western and northern parts of Denpasar. Kdr analysis of voltage-gated sodium channel (Vgsc) gene showed significant association to S989P and V1016G mutations linked to resistance phenotypes against 0.75% permethrin. Conversely, Ae. aegypti F1534C gene mutation did not result in any significant correlation to resistance development. CONCLUSIONS: Periodically surveillance of insecticide resistances in Ae. aegypti mosquitoes will help local public health authorities to set better goals and allow proper evaluation of on-going mosquito control strategies. Initial detection of insecticide resistance will contribute to conduct proper actions in delaying mosquito resistance development such as insecticide rotation or combination of compounds in order to prolong chemical efficacy in combating Ae. aegypti vectors in Indonesia.