Morphological and molecular characteristics of Plasmodium juxtanucleare in layer chicken from three districts of Yogyakarta, Indonesia.

Background and Aim: Blood parasite infections in poultry, such as Plasmodium, are a serious threat to the poultry industry due to their potential to cause economic losses. To date, there has been inadequate research on the morphological and molecular detection of the different Plasmodium species that infect poultry in Indonesia. Therefore, this study aimed to analyze the morphological and molecular characteristics of Plasmodium spp. and the several predisposing factors for Plasmodium infection in layer chickens from three districts of Yogyakarta, Indonesia. Materials and Methods: One hundred and five blood samples from layer chickens were collected from 13 farms located in three districts of Yogyakarta (Sleman, Bantul, and Kulon Progo) between September and November 2022. Blood samples were subjected to microscopic and polymerase chain reaction (PCR) analyses. Sequencing was performed using basic local alignment search tools to identify the nucleotide structure of cytochrome b. Phylogenetic analysis of Plasmodium was performed using the MEGA-X software. Results: Microscopic examination revealed that 17/105 positives (16.19%) were positive for blood parasite infection. Trophozoites, erythrocytic meronts, and microgametocytes of Plasmodium were found in blood samples. Based on the morphological examination, the species found in the samples was close to Plasmodium juxtanucleare. Polymerase chain reaction examination revealed that 21/60 samples were positive for Plasmodium (35%). The Plasmodium species identified from the sequenced samples were proven to be P. juxtanucleare. The P. juxtanucleare from Thailand was closely related to samples (99.64%-100%) with a genetic distance of 0%-1%. In addition, age, population, and cage type were not significantly associated with Plasmodium infection. Conclusion: Based on microscopic and PCR examinations, the Plasmodium species found in the three districts of Yogyakarta was P. juxtanucleare. The genetic distance between samples from the three districts of Yogyakarta was closely related (0%-1%) to P. juxtanucleare from Thailand and Japan. There was no correlation between Plasmodium infection and age, cage type, or population.

Anthelmintic activity of Carica pubescens aqueous seed extract and its effects on rumen fermentation and methane reduction in Indonesian thin-tailed sheep: An in vitro study.

Background and Aim: Seeds from Carica pubescens were observed to be abundant as waste. This waste contains active plant compounds whose utilization has not been studied in the livestock sector. This study was conducted to evaluate the potential of an aqueous seed extract of Carica pubescens (ASE) as an anthelmintic agent during rumen fermentation and methane reduction. Materials and Methods: Aqueous seed extract of Carica pubescens was prepared from C. pubescens cultivated in Wonosobo, Indonesia. Phytochemical analyses were performed to quantify the secondary metabolite content of ASE. In vitro adult worm mortality tests, scanning electron microscopy, and gas production tests were conducted to evaluate rumen characteristics, methane reduction, and the potential of ASE as an anthelmintic against Haemonchus contortus. Adult worms and ruminal fluid were collected from Indonesian thin-tailed sheep. Two-way analysis of variance followed by Tukey's test was performed using the Statistical Package for the Social Sciences® 21.0 software to detect significant differences. Results: In vitro study results showed that 1-5% ASE inhibited H. contortus after the 1st h of incubation, and 5% ASE suppressed 100% of adult H. contortus worms in the 5th h. Scanning electron microscopy analysis of ASE-treated worms ASE revealed damaged cuticle structures. ASE had no significant effect on pH, NH3, volatile fatty acid, acetate, propionate, butyrate, acetate: propionate, or microbial protein in rumen fluid (p > 0.05). The in vitro feed fermentation results showed that ASE significantly affected methane reduction. Conclusion: The inclusion of up to 5% ASE in sheep diets may serve as a potential alternative anthelmintic against H. contortus as well as a methane reduction agent, without deleterious effects on rumen fermentation.

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.

First Study of Ascaris lumbricoides from the Semiwild Population of the Sumatran Orangutan Pongo abelii in the Context of Morphological Description and Molecular Phylogeny.

There is little evidence that the already described and accepted taxa of ascarids (Ascaris lumbricoides, A. suum, and A. ovis) infecting individuals of taxonomically distant groups (hominids, pigs, sheep, goats, and dogs) can be genetically or morphologically distinguished. However, despite described morphological differences, e.g., due to intraspecific variation, these are insufficient for species determination and may indicate differences amongst ascarids because of cross infections, hybrid production, and specific adaptations to hosts. Herein, the results of a molecular and morphological analysis of ascarids parasitising Sumatran orangutans (Pongo abelii Lesson, 1827) in native populations are presented. The research took place in the Bukit Lawang area, Indonesia, in 2009. Throughout the year, fresh faecal samples were collected regularly from 24 orangutans, and all were examined for the presence of nematode adults. Only five adult worms from two orangutan females were found during regular collection. Using the integrative taxonomic approach, the nematodes found were identified as A. lumbricoides. The significance of the find and its rarity is documented by the fact that this is the first confirmed finding of adult ascarids from an original orangutan site (not from a zoo) in more than 130 years (including the long-term study spanning the last 20 years focusing on orangutan parasites and natural antiparasitic drugs). More accurate morphometric parameters and genetic differences for the identification of ascarids were established. These parameters will be helpful for other findings in great apes and will also be suitable for further and precise determination of this parasite. The details distinguishing between male and female specimens are also stated and well defined. A comprehensive evaluation of the situation of Ascaris species parasitising orangutans, including a comparison with previously described orangutan parasite (i.e., A. satyri-species inquirenda), is discussed.

Human social conditions predict the risk of exposure to zoonotic parasites in companion animals in East and Southeast Asia.

BACKGROUND: A recent dramatic surge in pet ownership has been observed across metropolitan areas in Asia. To date, there is a dearth of information on the risk associated with pet ownership for the transmission of parasites on a large scale in Asia, despite this continent giving rise to the largest burden of zoonotic infections worldwide. METHODS: We explored the nature and extent of zoonotic internal (endo-) and external (ecto-) parasites and arthropod-borne pathogens in 2381 client-owned dogs and cats living in metropolitan areas of eight countries in East and Southeast Asia using reliable diagnostic tests and then undertook extensive statistical analyses to define predictors of exposure to zoonotic pathogens. RESULTS: The estimated ORs for overall parasite infections are 1.35 [95% CIs 1.07;1.71] in young animals and 4.10 [1.50;11.2] in the animal group older than 15 years as compared with adult animals, 0.61 [0.48;0.77] in neutered animals as compared to unneutered animals, 0.36 [0.26;0.50] in animals living in urban areas as compared with rural areas, 1.14 [1.08;1.21] for each 1 °C increase of annual mean temperature which varies from 12.0 to 28.0 °C, and 0.86 [0.78;0.95] for each year of life expectancy which varies from 70.9 to 83.3 years. CONCLUSIONS: Here we highlight the influence of human life expectancy and the neutering status of the animals, which reflect increased living standards through access to education and human and veterinary health care, to be both strongly associated with exposure to zoonotic parasites. An integrated approach of local and international authorities to implement and manage educational programs will be crucial for the control of zoonotic infections of companion animals in Asia.

Parasites live on or inside animals or humans and can cause disease. Companion animals (pets) with parasites present a potential risk to the health of their owners, as certain kinds of parasites (known as zoonotic parasites) can affect both animal and human health. Here, we investigated whether human social conditions are associated with zoonotic parasite infections in companion animals in East and Southeast Asia. We found that higher human life expectancy and neutering of the companion animals were associated with fewer zoonotic parasite infections in the animals. These findings highlight the need for an enhanced commitment of local authorities to establish prevention campaigns, including education programs, against zoonotic pathogens. These measures will play a crucial role in alleviating the impact of these diseases in companion animals and humans in Asia.

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.

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.

Genetic characterization of nodular worm infections in Asian Apes.

Parasitic nematodes of Oesophagostomum spp., commonly known, as 'nodular worms' are emerging as the most widely distributed and prevalent zoonotic nematodes. Oesophagostomum infections are well documented in African non-human primates; however, the taxonomy, distribution and transmission of Oesophagostomum in Asian non-human primates are not adequately studied. To better understand which Oesophagostomum species infect Asian non-human primates and determine their phylogeny we analysed 55 faecal samples from 50 orangutan and 5 gibbon individuals from Borneo and Sumatra. Both microscopy and molecular results revealed that semi-wild animals had higher Oesophagostomum infection prevalence than free ranging animals. Based on sequence genotyping analysis targeting the Internal transcribed spacer 2 of rDNA, we report for the first time the presence of O. aculeatum in Sumatran apes. Population genetic analysis shows that there is significant genetic differentiation between Bornean and Sumatran O. aculeatum populations. Our results clearly reveal that O. aculeatum in free-ranging animals have a higher genetic variation than those in semi-wild animals, demonstrating that O. aculeatum is circulating naturally in wildlife and zoonotic transmission is possible. Further studies should be conducted to better understand the epidemiology and dynamics of Oesophagostomum transmission between humans, non-human primates and other wild species and livestock in Southeast Asia.

Genetic and parasitological identification of Trypanosoma evansi infecting cattle in South Sulawesi, Indonesia.

BACKGROUND AND AIM: Sulawesi is an Indonesian island located within the Wallacea region that contains a distinctive mix of Asian and Australasian species. This distinctiveness extends to parasites, including Trypanosoma evansi, the cause of surra. Surra has non-specific clinical signs such as anemia, anorexia, weight loss, drop in milk production, and reproductive disorders which cause economic losses. Due to the trade of livestock, surra has spread in Indonesia from one island to another. The aim of this study was to investigate the trypanosomes infecting cattle in South Sulawesi, using internal transcribed spacer (ITS2) ribosomal DNA (rDNA) sequencing. MATERIALS AND METHODS: A total of 100 whole blood samples were collected from cattle in Makassar, South Sulawesi Province, Indonesia. All samples were tested using conventional parasitological methods (CPT), namely, thin blood smear, buffy coat smears, and polymerase chain reaction (PCR) testing. Positive PCR results were sequenced and phylogenetically analyzed. RESULTS: Only one of the 100 samples was found to be positive with microscopic observation; however, PCR analysis revealed that 3% (3/100) of samples were positive. Sequencing identified the positive samples as T. evansi, China isolate (KU552344), with a homology of 99%. Two out of three sequences showed variations in ITS2 region. CONCLUSION: Based on CPT and molecular analysis, T. evansi isolates from infected cattle in South Sulawesi demonstrate genetic diversity of ITS2 sequences.

In vitro anthelmintic activity of aqueous and ethanol extracts of Paraserianthes falcataria bark waste against Haemonchus contortus obtained from a local slaughterhouse in Indonesia.

AIM: This study was conducted to determine the anthelmintic activity of aqueous and ethanol extracts of Paraserianthes falcataria bark against Haemonchus contortus. MATERIALS AND METHODS: Ethanol extract of bark (E.E.B.) waste and aqueous extract of bark (A.E.B.) waste of P. falcataria (at concentrations 0, 0.2, 0.4, 0.8, 1, 2.5, and 5%) and albendazole (2 mg/ml) as the positive control were placed in separate Petri dishes (50 mm). Twenty H. contortus worms were placed in Petri dishes and incubated at 37°C for 0.5, 1, 2, 3, 4, 5, 6, and 12 h. Mortality of each worm was ensured by pressing the body of the worm with a pair of tweezers and keeping it in lukewarm water for 5 min before declaring it dead. Mortality is defined as amount of death individuals and time of mortality of each worm was recorded. The parasites were then observed using scanning electron microscopy (SEM) at an accelerating voltage of 15 Kv. Statistical analysis was performed using SPSS 21.0 software, two-way ANOVA followed by Tukey's test to detect significant differences (p<0.05). The result was expressed as the mean ± standard deviation. RESULTS: The E.E.B. and A.E.B. of P. falcataria contained active compounds, such as tannin, alkaloid, flavonoid, saponin, steroid, and triterpenoid. E.E.B. had a higher content of phenol, while A.E.B. had a higher content of flavonoid. In this study, P. falcataria showed a significant effect (p=0.00) on H. contortus in vitro. E.E.B. (0.8%) was able to exterminate H. contortus completely after 6 h, more effective than A.E.B. (5%) while the positive control requires (2 mg/ml) after 2 h. SEM analysis of the worm treated with E.E.B. and A.E.B. showed damaged cuticle structure. CONCLUSION: The aqueous and ethanol extracts of P. falcataria bark waste demonstrated anthelmintic activity against H. contortus.

Molecular detection of pathogens in ticks and fleas collected from companion dogs and cats in East and Southeast Asia.

BACKGROUND: Ticks and fleas are considered amongst the most important arthropod vectors of medical and veterinary concern due to their ability to transmit pathogens to a range of animal species including dogs, cats and humans. By sharing a common environment with humans, companion animal-associated parasitic arthropods may potentially transmit zoonotic vector-borne pathogens (VBPs). This study aimed to molecularly detect pathogens from ticks and fleas from companion dogs and cats in East and Southeast Asia. METHODS: A total of 392 ticks and 248 fleas were collected from 401 infested animals (i.e. 271 dogs and 130 cats) from China, Taiwan, Indonesia, Malaysia, Singapore, Thailand, the Philippines and Vietnam, and molecularly screened for the presence of pathogens. Ticks were tested for Rickettsia spp., Anaplasma spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. while fleas were screened for the presence of Rickettsia spp. and Bartonella spp. RESULT: Of the 392 ticks tested, 37 (9.4%) scored positive for at least one pathogen with Hepatozoon canis being the most prevalent (5.4%), followed by Ehrlichia canis (1.8%), Babesia vogeli (1%), Anaplasma platys (0.8%) and Rickettsia spp. (1%) [including Rickettsia sp. (0.5%), Rickettsia asembonensis (0.3%) and Rickettsia felis (0.3%)]. Out of 248 fleas tested, 106 (42.7%) were harboring at least one pathogen with R. felis being the most common (19.4%), followed by Bartonella spp. (16.5%), Rickettsia asembonensis (10.9%) and "Candidatus Rickettsia senegalensis" (0.4%). Furthermore, 35 Rhipicephalus sanguineus ticks were subjected to phylogenetic analysis, of which 34 ticks belonged to the tropical and only one belonged to the temperate lineage (Rh. sanguineus (sensu stricto)). CONCLUSION: Our data reveals the circulation of different VBPs in ticks and fleas of dogs and cats from Asia, including zoonotic agents, which may represent a potential risk to animal and human health.

Zoonotic Vectorborne Pathogens and Ectoparasites of Dogs and Cats in Eastern and Southeast Asia.

To provide data that can be used to inform treatment and prevention strategies for zoonotic pathogens in animal and human populations, we assessed the occurrence of zoonotic pathogens and their vectors on 2,381 client-owned dogs and cats living in metropolitan areas of 8 countries in eastern and Southeast Asia during 2017-2018. Overall exposure to ectoparasites was 42.4% in dogs and 31.3% in cats. Our data cover a wide geographic distribution of several pathogens, including Leishmania infantum and zoonotic species of filariae, and of animals infested with arthropods known to be vectors of zoonotic pathogens. Because dogs and cats share a common environment with humans, they are likely to be key reservoirs of pathogens that infect persons in the same environment. These results will help epidemiologists and policy makers provide tailored recommendations for future surveillance and prevention strategies.

Entamoeba histolytica infections in wild and semi-wild orangutans in Sumatra and Kalimantan.

Key to the success of orangutan conservation management practices is the prevention of the introduction of infectious diseases to the remaining populations. Previous reports of Entamoeba spp. positive orangutans are of concern as Entamoeba spp. infection has been linked to morbidity and mortality in primates. It remains to be determined if the Entamoeba species infecting orangutans is the pathogenic Entamoeba histolytica. Orangutan fecal samples have been collected from orangutans from sites in Sumatra (Bukit Lawang, Ketambe, and Suaq, 241 samples from 64 individuals), and two sites in Kalimantan (Sebangau and Tuanan, 129 samples from 39 individuals). All samples were from wild orangutans except for a proportion from Sumatra which were from semi-wild (108 samples, 10 individuals). E. histolytica-specific nested PCR assays were carried out on the fecal samples. A total of 36 samples from 17 individuals tested positive for E. histolytica. When compared with published sequences using NCBI BLAST the E. histolytica positive samples showed a 98-99% concordance. The majority (76%, n = 36) of the positive isolates came from semi-wild orangutans in Bukit Lawang. This study supports the growing body of evidence that contact with humans is an important risk factor for infection of wild primates with E. histolytica.

Morphological and molecular identification of Pfenderius heterocaeca (Trematode: Paramphistomoidea) from Sumatran elephant (Elephas maximus sumatranus).

BACKGROUND AND AIM: Paramphistomiasis is common in tropical countries such as Indonesia and affects livestock and various endemic wild animals such as Sumatran elephants. However, the specific species of paramphistomoid worm that causes paramphistomiasis are rarely reported. The study aims at identifying paramphistomoid worm that infects Sumatran elephants. MATERIALS AND METHODS: Flukes were collected from the feces of five semi-captive Sumatran elephants that lived at Tegal Yoso Elephant Response Unit in Way Kambas National Park, in 2018, after treatment of oxyclozanide 1 g at the dose of approximately 5-8 mg/kg of body weight. Eight paramphistomoid worms were flattened and stained in Semichon's carmine for morphological identification, and five other worms were used for molecular identification at second internal transcribed spacer (ITS-2) of ribosomal deoxyribonucleic acid sequence. RESULTS: Forty-five flukes were collected from five Sumatran elephants in Lampung, Indonesia. Eight paramphistomoid worms were morphologically identified as Pfenderius heterocaeca and five isolates did not show any variation in ITS-2. Phylogenetic analysis showed that there was a close genetic relationship between our sample and Chiorchis fabaceus that had a family similar to the samples. CONCLUSION: Based on the morphological and molecular characteristics, the paramphistomoids found in Sumatran elephant on Way Kambas National Park are P. heterocaeca.

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.

Molecular characterization of highly pathogenic Eimeria species among beef cattle on Java Island, Indonesia.

Gastrointestinal parasites including Eimeria spp. are known to affect domestic animal productivity causing watery or lethal bloody diarrhea. However, there are few reports on the detailed distribution of bovine Eimeria spp. in cattle, particularly in developing tropical and sub-tropical areas. Using a total of 289 fecal samples collected from beef cattle on Java Island, one of the five main islands of Indonesia, fecal examinations by the Whitlock and sugar flotation methods and molecular surveys were conducted to reveal the prevalence of 6 Eimeria spp. As a result of morphological screening using Whitlock methods and sugar flotation, Eimeria spp. prevalences of 9.4% and 52.3% were confirmed, respectively. The prevalence was higher in younger cattle [under 1 year (63.9%), 1-2 years (75.0%) and more than in 2 year old cattle (42.3%)]. The prevalences of identified species were as follows: 10.4% for E. bovis, 2.8% for E. ellipsoidalis, 2.1% for E. alabamensis, 1.4% for E. zuernii, 1.1% for E. auburnensis, and 0.4% for E. cylindrica. Moreover, prevalences of 12.8% for Strongyloides papillosus, 7.3% for Trichuris globulosa, and 0.3% for Capillaria bovis were detected. Although the average number of oocysts per gram of feces was <100 among the positive samples, and cases of heavy infection were limited, the endemicity of these pathogenic Eimeria species among farms in Indonesia should be noted.

Morphology and morphometry of adult nematodes on Sumatran elephants (Elephas maximus sumatranus) in Way Kambas National Park area, Indonesia.

BACKGROUND AND AIM: Worms from nematodes are the most numerous and the most detrimental in elephants. Most adult worms are located in the digestive tract. Nematode infection is at higher risk in young elephants, which caused several cases such as anemia, hypoalbuminemia, enteritis, and even death. This study aimed to determine the morphology and morphometry of adult nematodes on Sumatran elephants in Way Kambas National Park area. MATERIALS AND METHODS: Nematode samples were obtained from Sumatran elephants' feces (Elephas maximus sumatranus) in Way Kambas National Park, Lampung Province, after being given Kalbazen® containing albendazole 1000 mg at a dose of 10 mg/kg by the veterinarian in charge of the National Park area. For the morphological and morphometric examinations, we used an Olympus BX 51 microscope equipped with Olympus DP 12 camera and were conducted at the Parasitology Laboratory, Faculty of Veterinary Medicine, Universitas Gadjah Mada. The scanning electron microscopic (SEM) analysis was carried out at the Biology Research Center of the Indonesian Institute of Sciences (Lembaga Ilmu Pengetahuan Indonesia). RESULTS: The results of macroscopic observations of the obtained nematodes showed that the nematodes which were found have the characteristics of round, slim, and white color. The size of a female worm was larger than a male worm. Microscopic examination in four anterior papillae indicated that the dorsal lobe in the copulatory bursa was longer than lateral lobe. The result of inspection with the SEM showed a leaf crown consisting of 10 elements, a pair of amphids laterally, and two pairs of papilla in a submedian region. CONCLUSION: Based on our morphology and morphometry examinations of adult nematodes in Sumatran elephant (E. maximus sumatranus) in Way Kambas National Park area, the adult nematodes which were found are species of Quilonia travancra.

Prevalence of gastrointestinal worms in Wonosobo and thin-tailed sheep on the slope of Mount Sumbing, Central Java, Indonesia.

AIM: This study was conducted to determine the prevalence of gastrointestinal (GI) worms in Wonosobo and thin-tailed sheep from the slope of Mount Sumbing. MATERIALS AND METHODS: Fecal samples (n=305) were collected directly from the rectum of Wonosobo and thin-tailed sheep during the dry and rainy seasons in Wonosobo Regency, Central Java Province, Indonesia. The presence of GI helminth eggs in the fecal samples was assessed using the modified McMaster egg counting technique. The identification of the eggs or oocysts was done on the basis of their morphology and size. RESULTS: The highest prevalence of GI worms was observed in male thin-tailed sheep (76.47%) during the rainy season, whereas the lowest prevalence was observed in female Wonosobo sheep (47.36%) during the dry season. The types of GI nematodes observed in these two types of sheep were Haemonchus contortus, Ostertagia spp., Trichostrongylus spp., Bunostomumspp., Trichuris spp., and Moniezia spp. The GI worms with the highest prevalence were of Haemonchus spp. and were observed in male thin-tailed sheep. The prevalences of the two types of sheep assessed at an altitude of 1150 m above sea level were higher than those observed at the altitude of 1586 m. The prevalence of clinical and sub-clinical parasites infestation in Wonosobo and thin-tailed sheep in Kwadungan village was significantly different (p<0.05). CONCLUSION: This study showed that two types of local sheep on the slope of Mount Sumbing are infected with various GI worms during the dry and rainy seasons. The highest prevalence of GI worms was found in thin-tailed sheep at an altitude of 1150 m above sea level during the rainy season, with H. contortus being the most prevalent GI parasites.