Molecular identification of new Trypanosoma evansi type non-A/B isolates from buffaloes and cattle in Indonesia.

Trypanosoma evansi is reportedly divided into two genotypes: types A and B. The type B is uncommon and reportedly limited to Africa: Kenya Sudan, and Ethiopia. In contrast, type A has been widely reported in Africa, South America, and Asia. However, Trypanosoma evansi type non-A/B has never been reported. Therefore, this study aims to determine the species and genotype of the Trypanozoon subgenus using a robust identification algorithm. Forty-three trypanosoma isolates from Indonesia were identified as Trypanosoma evansi using a molecular identification algorithm. Further identification showed that 39 isolates were type A and 4 isolates were possibly non-A/B types. The PML, AMN-SB1, and STENT3 isolates were likely non-A/B type Trypanosoma evansi isolated from buffalo, while the PDE isolates were isolated from cattle. Cladistic analysis revealed that Indonesian Trypanosoma evansi was divided into seven clusters based on the gRNA-kDNA minicircle gene. Clusters 6 and 7 are each divided into two sub-clusters. The areas with the highest genetic diversity are the provinces of Banten, Central Java (included Yogyakarta), and East Nusa Tenggara. The Central Java (including Yogyakarta) and East Nusa Tenggara provinces, each have four sub-clusters, while Banten has three.

Tissue cysts and serological detection toxoplasmosis among wild rats from Surabaya, East Java, Indonesia.

Background: The protozoan Toxoplasma gondii is the source of zoonosis toxoplasmosis and causes public health problems throughout the world. Environmental contamination by oocysts excreted by cats as definitive hosts affects the spread of this disease. Wild rats as rodents can be used as an indicator of environmental contamination by oocysts, considering that rats have a habit of living in dirty environments and can be infected by oocysts from the environment. Aim: This study aims to detect toxoplasmosis from tissue cysts and serological tests in wild rats as an indicator of environmental contamination in Surabaya. Methods: A total of 100 wild rats collected from Surabaya were collected in five areas (West, East, Central, North, and South of Surabaya) obtained from three trapping locations: housing, dense settlements, and markets. All samples were examined microscopically for parasitological tests through the brain tissue samples, and the serum was examined using the toxoplasma modified agglutination test to detect the presence of IgG and Immunoglobulin M (IgM). Results: This research used 100 wild rat samples, 77 Rattus tanezumi and 33 Rattus norvegicus, with evidence of 31% in serology and active infection with 19% tissue cyst. The results showed that the seroprevalence of T. gondii in wild rats was 31% (30% for IgG and 1% for IgM). Tissue cysts in the rat brain samples tested were 19% (19/100). The IgG prevalence rate in female rats was 25% (8/32), while for males, it was 32.3% (22/68). The highest seropositive IgG from densely populated settlements was 50%, markets were 25.8%, and housing was 12.1%. The highest seropositive IgM from densely populated settlements was 2.8%. Population density and the presence of cats are factors supporting the high seropositive rate at the trapping location. Conclusion: This study revealed that there has been toxoplasmosis contamination in Surabaya with evidence of 31% in serology and active infection with 19% tissue cyst. It is necessary for controlling with surveillance in cats to prevent transmission in humans.

Serological survey of immunoglobulin G from Toxoplasma gondii infection in dairy goats in East Java, Indonesia.

Background and Aim: Toxoplasma gondii infection is a significant issue of veterinary public health because it is potentially transmitted through goat milk. Therefore, the use of control measures and routine monitoring of toxoplasmosis in dairy goats is necessary. Serological analysis using antibodies can detect T. gondii infection. This study aimed to conduct an epidemiological study of T. gondii in dairy goats using antibody detection and risk factor identification. Materials and Methods: This was a cross-sectional study. We performed a serological analysis of T. gondii infection in dairy goats to evaluate the prevalence of toxoplasmosis. Random sampling was performed, including 132 lactating dairy goats. Toxoplasma-modified agglutination test was used as a serological test for immunoglobulin G with a sensitivity of 98.55%, specificity of 86.21%, and accuracy of 94.9%. A structured questionnaire was used to collect risk factor data, which were analyzed using the Chi-square and Fisher's exact tests. The statistical package for the social sciences v. 21 was used for statistical analyses. Results: The seroprevalence of T. gondii in Malang and Lumajang Regency was 100% and 90.7%, respectively. A significant difference in prevalence of T. gondii was observed between the two districts. Livestock management practices that significantly influenced T. gondii seroprevalence included water sources (p < 0.05; relative risk [RR] = 1.151; 95% confidence interval [CI]: 1.044-1.269). Farmers' characteristics that significantly influenced T. gondii seroprevalence included education (p < 0.05; RR = 1.125; 95% CI: 1.037-1.221), main occupation (p < 0.05; RR = 1.118; 95% CI: 1.035-1.207), and position in the organization of dairy goats farmers (p < 0.05; RR = 1.141; 95% CI: 1.022-1.274). Conclusion: In East Java, the prevalence of T. gondii in dairy goats is high. This study provides detailed information regarding risk factors associated with T. gondii seroprevalence in dairy goats in East Java, Indonesia.

Designing one-step reverse transcriptase loop-mediated isothermal amplification for serotype O foot-and-mouth disease virus detection during the 2022 outbreak in East Java, Indonesia.

Background and Aim: Various methods can detect foot-and-mouth disease (FMD) in cows, but they necessitate resources, time, costs, laboratory facilities, and specific clinical specimen submission, often leading to FMD virus (FMDV) diagnosis delays. The 2022 FMD outbreak in East Java, Indonesia, highlighted the need for an easy, inexpensive, rapid, and accurate detection approach. This study aims to devise a one-step reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) technique and phylogenetic analysis to detect the serotype O FMDV outbreak in East Java. Materials and Methods: Swab samples were collected from the foot vesicles, nasal secretions, and saliva of five suspected FMDV-infected cows in East Java between June and July 2022. The RT-LAMP design used hydroxy naphthol blue dye or SYBR Green I dye, with confirmatory analysis through reverse transcriptase polymerase chain reaction (RT-PCR) targeting 249 base pairs. PCR products underwent purification, sequencing, and nucleotide alignment, followed by phylogenetic analysis. Results: The RT-LAMP method using hydroxy naphthol blue dye displayed a positive reaction through a color shift from purple to blue in the tube. Naked-eye observation in standard light or ultraviolet (UV) light at 365 nm, with SYBR Green I stain, also revealed color change. Specifically, using SYBR Green I dye, UV light at 365 nm revealed a color shift from yellow to green, signifying a positive reaction. Nucleotide alignment revealed mutations and deletion at the 15th sequence in the JT-INDO-K3 isolate from the East Java FMDV outbreak. Despite differing branches, the phylogenetic tree placed it in the same cluster as serotype O FMDV from Malaysia and Mongolia. Conclusion: JT-INDO-K3 exhibited distinctions from Indonesian serotype O FMDV isolates and those documented in GenBank. Then, the RT-LAMP method used in this study has a detection limit 10 times higher latter than the conventional RT-PCR limit, without any cross-reactivity among strains.

A parasitological survey and the molecular detection of Entamoeba species in pigs of East Java, Indonesia.

Background and Aim: In several countries, two Entamoeba porcine species, Entamoeba suis and Entamoeba polecki (subtype 1 and 3), have been detected using molecular methods and identified pathogenicity associated with enteritis. However, globally, Entamoeba infection prevalence in pigs is extremely limited. This study aimed to coprologically and genetically examine pig parasites to estimate prevalence of Entamoeba in three pig farms in East Java, Indonesia. Materials and Methods: Hundred porcine fecal samples (Landrace) were collected from three East Javan farms in well-known swine industry regions. Fecal samples were examined under a microscope after sugar-flotation centrifugation, and molecular species and subtype identification were performed using polymerase chain reaction (PCR) and primer pairs targeting small-subunit ribosomal RNA. Results: Microscopy examinations identified parasites in 89/100 fecal samples; Entamoeba spp. cysts were the most frequent in these samples. Polymerase chain reaction showed that 58 samples were comprised of mixed Entamoeba suis and Entamoeba polecki, 22 E. suis alone, and nine E. polecki alone infections. Epolec F6-Epolec R6 primers successfully amplified E. polecki ST1-4 subtypes, while Epolecki 1-Epolecki 2 amplified only the E. polecki ST1 subtype. Entamoeba polecki ST1-specific primers successfully detected the ST1 subtype in 19/67 E. polecki positive samples. Conclusion: Entamoeba spp. prevalence in Indonesian pigs was previously shown to be high. On coprological examination of East Javan pigs, we detected high Entamoeba spp. levels, in which we genetically identified as E. suis (80.0%), E. polecki (67.0%), and E. polecki ST1 (19%).

Aeromonas hydrophila induction method in adult zebrafish (Danio rerio) as animal infection models.

Background and Aim: Zebrafish are frequently used as model organisms in scientific research as their genes mirror those of humans. Aeromonas hydrophila bacteria can infect humans and animals, mainly fish. This study aimed to identify the concentration and route of A. hydrophila infection in adult zebrafish. Zebrafish had been used as a challenge test by analyzing their hematological profiles, blood glucose levels, and survival rates. Materials and Methods: Induction of cell supernatant free (CSF) from A. hydrophila bacteria in adult zebrafish was carried out via bath immersion (BI), intraperitoneal injection (IPI), intramuscular injection (IMI), and healthy zebrafish as a control (C). The bacterial concentrations were 107, 109, and 1011 colony-forming units (CFU)/mL. At 24 h post-infection, the outcomes of infection were evaluated based on survival rates, hematological profiles, and blood glucose levels. A one-way analysis of variance with a confidence level of 95% was employed to examine the data. Results: In the BI, IPI, and IMI treatment groups, the survival rate of the fish reached a peak of 100%, 22%-100%, and 16%-63%, respectively, compared with the injection technique. In the IMI2 group, a 109 CFU/mL bacterial concentration was determined to correspond to the lethal dosage 50. All infection groups had lower erythrocyte and hemoglobin counts but higher leukocyte counts than the control group. The blood sugar levels of the healthy and infected groups were not significantly different. Conclusion: The route of A. hydrophila infection through Intramuscular injection with a concentration of 109 CFU/mL indicated a high performance compared to other techniques. This method could be developed as a reproducible challenge test.

Methanol extract of Black soldier fly (Hermetia illucens) prepupae against Aeromonas and Staphylococcus aureus bacteria in vitro and in silico.

Background: Staphylococcus and Aeromonas bacteria are pathogens in humans and animals. The therapy disrupts the virulence structure of the bacteria, resulting in bacterial death. Currently, chemical drugs have resulted in many resistant bacteria, so it is necessary to find alternative natural materials that are not toxic and do not quickly induce resistance. Aims: This study aimed to analyze the potential of methanol extract from Black soldier fly (BSF) prepupae as an antibacterial agent against Staphylococcus aureus and Aeromonas through in silico and in vitro tests. Methods: The BSF prepupae methanol extract was analyzed for protein and fatty acid contents. Disc diffusion method, minimal inhibitory concentration, and minimum bactericidal concentration test were used for in vitro tests against Staphylococcis and Aeromonas. Molecular docking of the active ingredients (defensin, chitin, and chitosan as well as fatty acids) in BSF was downloaded from the NCBI database and docked by the Hex Cuda version 8.0 program with Correlation type parameters Shape + Electro and Grid Dimension version 0.6. Docking results were analyzed using the Discovery Studio program version 21.1.1. Results: The highest fatty acid contents in the extract were palmitic acid and myristic acid. Methanol extract from BSF prepupae acted as a bactericidal agent against S. aureus at a concentration of 320 mg/ml, in contrast to Aeromonas, which still showed bacterial growth. The results of the in silico test showed that defensin-aerolysin and defensin-hemolysin was bound to the same active site area. However, the amount of binding energy produced by 69-Defensin-83-aerolysin was higher than all defensin types in BSF against Aeromonas. Chitin and chitosan showed a bond on the active site of aerolysin and hemolysin, but chitosan had a stronger bond than chitin. In silico study also showed the strongest binding affinity of BSF fatty acids to isoleucyl-tRNA synthetase of S. aureus. Conclusion: The study showed that methanol extract from BSF prepupae had potential capability as an antibacterial agent against S. aureus than Aeromonas in vitro and in silico.

Comparison of ITS-1 and TBR-1/2 primer sensitivity for the detection of Trypanosoma evansi local isolates in experimental rats using a polymerase chain reaction.

Background and Aim: Surra is caused by Trypanosoma evansi. The detection method using conventional parasitological tests has not always shown positive results in blood parasite detection, although the livestock has presented with clinical signs. Therefore, a fast and accurate diagnosis is necessary to prevent the disease predominately in field isolates. This study aimed to investigate the sensitivity of molecular detection method using two different specific primers, namely, Internal Transcribed Spacer 1 (ITS-1) and Trypanosoma brucei repeat 1/2 (TBR-1/2) against T. evansi field isolates from Banten Province, Indonesia. Materials and Methods: The isolates of T. evansi used in this study were collected from Banten Province and cultured and preserved by the National Research Center for Veterinary Science, Indonesia. Eighteen experimental rats were divided into three equal groups, which were categorized as control, 1 × 101, and 1 × 104 infective doses. The isolates were injected into all experimental albino rats intraperitoneally. All samples were tested using conventional blood smear, card agglutination test (CATT), and polymerase chain reaction (PCR) method. Results: The results of the CATT examination in all treatments showed negative results. However, PCR results showed that two different primers, namely, ITS-1 and TBR-1/2 had been successfully detected T. evansi from infected experimental rats, proven by positive PCR band appeared in 480 base pairs (bp) and 164 bp, respectively. Conclusion: Based on the molecular diagnostic test using PCR method, TBR-1/2 primer is more sensitive to detect T. evansi compared to ITS-1 primer. The present finding provides preliminary data for studying the efficiency of different primers if practically applied as a standard diagnostic test for trypanosomiasis, especially in Indonesian livestock.

Occurrence and biodiversity of Eimeria spp. (Apicomplexa: Eimeriidae) in Madura cattle reared on Kamal Subdistrict, Madura Island, Indonesia.

Background and Aim: In Indonesia, Madura cattle are native breeds that are expected to contribute to the improvement of regional meat self-sufficiency. Eimeria spp. are protozoans that are commonly found in ruminants. This study aimed to identify the occurrence and diversity of Eimeria spp. in Madura cattle. Materials and Methods: In this study, fresh fecal samples were collected from 100 cattle in Kamal Subdistrict, Bangkalan District, Madura Island, Indonesia. Morphological detection was performed using a light microscope, and molecular identification was performed using a polymerase chain reaction. DNA amplification was conducted using various species-specific primers for Eimeria bovis, Eimeria zuernii, Eimeria auburnensis, Eimeria alabamensis, Eimeria ellipsoidalis, and Eimeria cylindrica. Results: The results obtained 21% (21/100) of Eimeria spp. based on morphological detection. A total of 15 positive samples with 500-25,000/mL oocysts were selected for DNA extraction and amplification, resulting in 12 positive samples. Four Eimeria spp. were obtained based on molecular identification: E. bovis, E. zuernii, E. auburnensis, and E. cylindrica. Conclusion: Four species of Eimeria namely E. bovis, E. zuernii, E. auburnensis, and E. cylindrica were identified from fecal sample of Madura cattle using PCR method in this study. Further comprehensive studies are required to investigate the pathogenicity of Eimeria spp. in Madura cattle. Therefore, improved and integrated management practices should be strengthened by local governments to prevent pathogenic diseases and increase national livestock productivity in Indonesia.

Clove Flower Extract (Syzygium aromaticum) Has Anticancer Potential Effect Analyzed by Molecular Docking and Brine Shrimp Lethality Test (BSLT).

Various anticancer medications have been discovered due to advances in the health care industry, but they have undesirable side effects. On the other hand, anticancer drugs derived from natural sources have low side effects, making them excellent for cancer therapy. This study aims to evaluate the effect of clove flower extract (Syzygium aromaticum) as a potential anticancer agent by determining grid-score values using molecular docking and LC50 values using the brine shrimp lethality test (BSLT) technique. As animal models, three hundred larvae of Artemia salina leach were divided into six groups. Each group has ten larvae that have undergone five replications. The clove flower extract concentration in the treatment media was 50 ppm (T1), 250 ppm (T2), 500 ppm (T3), 750 ppm (T4), 1000 ppm (T5), and 0 ppm (seawater) as the control. The probit analysis of Artemia Salina leach mortality percentage data. The results indicated that the clove flower extract (Syzygium aromaticum) is harmful to larvae with LC50 values of 227,1 g/ml or in the equation y = 2,8636 x - 1,7466 with an R2 value of 0.9062. According to molecular docking, eugenol acetate (grid-score -42.120834) has a close relationship with the cognate enzyme nitric oxide synthase (3E7G) based on its proximity to the grid score value (grid-score -61.271812). Therefore, clove flower extract has the potential to act as an anticancer medication. Based on the grid-score proximity, eugenol acetate is close to the homologous enzyme nitric oxide synthase (3E7G). Inhibition of nitric oxide synthase also shows a reduction in cancer cell proliferation.

Seroprevalence of Toxoplasma gondii and associated alterations in hematology and serum biochemistry of one-humped camels (Camelus dromedarius) in Pakistan.

Background and Aim: Toxoplasma gondii is an intracellular protozoan that infects humans and animals. This study aimed to estimate the seroprevalence of T. gondii and the associated alterations in hematology and serum biochemistry of one-humped camels (Camelus dromedarius) in Mianwali district, Pakistan. Materials and Methods: A total of 350 blood samples were obtained from male and female camels of different ages (≤3 years old, 4-6 years old, and ≥7 years old). To validate T. gondii antibodies, the collected samples were subjected to indirect enzyme-linked immunosorbent assay using purified recombinant micronemal protein 3 as an antibody catching antigen. Results: The prevalence of T. gondii was 50.2% higher in male camels than in female camels (16.5%) (p<0.001). Furthermore, the prevalence of T. gondii in camels was directly proportional to age (p<0.001). It was 63.33% (57/90) in camels of ≥7 years of age, 32.54% in 4-6 years old age group, and 23.08% in ≤3 years old age group. The hematological analysis of infected camels revealed a significant increase in the values of glucocorticoid-remediable aldosteronism, lymphocyte percentage, monocyte percentage (MONO%), corpuscular hemoglobin (MCH), and procalcitonin. Furthermore, substantially higher levels of liver enzymes alanine aminotransferase, aspartate aminotransferase, and the macro-mineral potassium were found in the serum of T. gondii-infected camels. Conclusion: The seropositivity of T. gondii is directly associated with the age and sex of camels, which may be considered as potential risk factors. Furthermore, T. gondii infection directly impacts the hemato-biochemistry of infected camels.

Coproparasitological examinations and molecular determination of Eimeria species in Madura cattle reared on Madura Island, Indonesia.

Madura cattle, which are native to Indonesia and mainly kept on Madura Island, East Java, are expected to contribute to improving the regional meat self-sufficiency. Eimeria spp. are the most pathogenic protozoans among gastrointestinal parasites in livestock but no molecular surveys of Eimeria spp. in Madura cattle have been conducted to date. In this study, a total of 183 fecal samples were collected from Madura cattle and 60 (32.8%) were positive for parasites of protozoans and nematodes by the sugar floatation method. Among the samples with parasites, Eimeria spp. oocysts were detected in 50 samples (27.3%) with an average OPG value of 1686.1. Eimeria spp. were successfully identified to the species level in 26 samples with Eimeria bovis being the most prevalent, followed by E. zuernii and E. aubrunensis. A total of 21 samples showed mixed infection of more than two species of Eimeria. E. bovis and E. zuernii have been recognized as having high virulency and, thus, these parasites are potential sources of severe coccidiosis and the cause of infections in other cattle. Although additional studies are warranted, these results can be helpful for improving the management and productivity of Madura cattle.

Probiotic Pediococcus pentosaceus ABY 118 to Modulation of ChIFN-γ and ChIL-10 in Broilers Infected by Eimeria tenella Oocyst.

Eimeria causes coccidiosis, which has long been recognized as a disease in chickens that significantly affects the economy. The global chicken population continues to grow, and its contribution to food security increases, making it increasingly important to produce chicken meat that is safe for human and health. This study aims to prove Pediococcus pentosaceus ABY 118 to modulation of ChIFN-γ and ChIL-10 in chickens infected with E. tenella oocysts. This study used 100 of day-old chickens (DOC), randomly divided into 5 treatments; each treatment consists of 20 chickens. The treatments was as follows: P0 (-): negative control; P0 (+): positive control; P1: monensin; P2: probiotic 1.5 × 108 CFU/ml; and P3: probiotic 3.0 × 108 CFU/ml. At the age of 20 days, Eimeria tenella (E. tenella) oocysts were inoculated orally at a dose of 1 × 104. The probiotic P. pentosaceus ABY 118 was given orally through drinking water from DOC to 35 days. Monensin was given orally through feed from the age of 14-26 days. The results of statistical analysis showed that there was a significant difference (P < 0.05) between treatments on ChIFN-γ and ChIL-10 at 6 and 8 days postinfected with E. tenella oocysts. Based on the results of this study, it can be concluded that the use of P. pentosaceus ABY 118 isolates at a dose of 1.5 × 108 CFU/ml and 3.0 × 108 CFU/ml per liter of drinking water can increase health by stimulation of ChIFN-γ and ChIL-10 in broiler infected with E. tenella oocyst.

Zoonotic and other gastrointestinal parasites in cats in Lumajang, East Java, Indonesia.

Relationship between humans and cats has negative impact associates with zoonotic diseases. It is the reason why studies on the prevalence of gastrointestinal (GI) parasites in cats are important. Some of zoonotic GI parasites in cats are Toxocara spp, Ancylostoma sp, and Toxoplasma gondii. The current study was conducted to investigate the prevalence of GI parasites in owned and stray cats in Lumajang East Java Indonesia. One hundred and twenty fecal samples were collected from owned and stray cats on November 2018 to January 2019. The samples were examined by direct smears, sedimentation and flotation techniques. Identification of parasites was determined based on the morphology of worm eggs and protozoan cysts. The results showed that gastrointestinal parasites were found in 68.33% (82/120) examined samples, respectively, 48.33% (29/60) and 88.33% (53/60) from owned cats and stray cats. We found 7 genera of parasites, 4 genera of worm eggs and 2 genera protozoan oocyst. The egg worm were Toxocara cati (40 %), Toxocara leonina. (10.33%), Ancylostoma sp. (18.33%), Diphylobothrium sp. (3.33%) and Dipylidium caninum (1.67%). The protozoan oocyst were Isospora felis (27.5%), Isospora rivolta (13.33%) and Eimeria spp. (8.33%). Toxocara cati, Ancylostoma sp. (hookworm), Diphylobothrium sp. and Dipylidium caninum were zoonotic parasites. Rate infection in younger and older cat were no significant difference. One cat can be infected one or more parasite. To conclude, the prevalence of zoonotic GI parasites both in owned and stray cats were high. It is necessary to plan a program to control this zoonotic parasites.

Blastocystis spp. subtype 10 infected beef cattle in Kamal and Socah, Bangkalan, Madura, Indonesia.

BACKGROUND AND AIM: Blastocystis spp. is a gastrointestinal parasite that can infect both humans and animals and has the potential to become a zoonotic parasite. This study analyzed a subtype (ST) of Blastocystis spp. that had infected beef cattle in Kamal and Socah, Bangkalan, Madura, Indonesia. MATERIALS AND METHODS: Fresh stool samples were collected from 108 beef cattle at Kamal and Socah, Bangkalan, Madura, Indonesia. Blastocystis spp. were detected both morphologically and genetically based on the 18S rRNA gene. The morphology of Blastocystis spp. from the stool samples and cultured samples were observed under a light microscope. Blastocystis spp. from 20 positive cultures were amplified through polymerase chain reaction, and the resultant sequences were identified by ST. RESULTS: One hundred and eight (100%) fecal samples from the fresh or cultured stools were positive morphologically for Blastocystis spp. Molecularly, all 20 of the samples selected for DNA analysis were found to be Blastocystis spp. ST 10. CONCLUSION: Based on morphological and molecular detection, the prevalence of Blastocystis spp. infection in beef cattle within Kamal and Socah, Bangkalan, Madura, Indonesia, was high. About 100% were non-zoonotic parasites. This was the first report of Blastocystis spp. ST 10 found in infected beef cattle in Kamal and Socah, Bangkalan, Madura, Indonesia.

Prevalence and diversity of gastrointestinal protozoa in Madura cattle at Bangkalan Regency, East Java, Indonesia.

AIM: This study aimed to describe the gastrointestinal protozoa in Madura cattle at Bangkalan Regency, East Java, Indonesia. MATERIALS AND METHODS: A total of 500 samples of Madura cattle feces were collected from 10 districts at Bangkalan Regency. Those ten districts represent the lowland and upland areas, and each district was represented by one village. The collected feces were examined using native, sedimentation, and floating methods. The species identification was determined by their morphology. RESULTS: There were 357 (71.4%) samples positively infected with protozoan. The highest rate of sample with protozoan infection was at Kamal District (88.23%), and Bangkalan District (52.83%) was the lowest one. There were six species of protozoa that infected gastrointestinal tract; those are Eimeria spp., Balantidium spp., Isospora spp., Blastocystis spp., Entamoeba spp., and Cryptosporidium spp. The highest number of protozoa found in this research was Eimeria (53.42%) followed by Blastocystis (14.43%). In this study, we found that 295 samples (58.76%) infected by one kind of protozoa, 53 samples (10.56%) infected by two kinds of protozoa, and 11 samples (2.19%) infected by three kinds of protozoa. In addition, there were 65.54% of bulls infected with protozoa, considerably lower than cows (72.97%). Cattle aged 6 months-2 years old (73.39%) and >2 years old (71.25%) are known more prone to protozoan infections than cattle aged <6 months (66.15%). CONCLUSION: The present study revealed that protozoan infection of cattle is common in Bangkalan Regency. Studies focused on determining that the prevalence of protozoan, risk factors for the parasitism, and the geographic distribution are needed and will be effective guide for prevention and control measures.

Exploration of Sarcoptes scabiei Antigenic Protein Which Play Roles in Scabies Pathogenesis in Goats and Rabbits.

BACKGROUND: Scabies or mange is an infectious skin disease caused by the mite Sarcoptes scabiei. This skin disease affects various livestock such as goats, sheep, swine, cattle, other animals like dogs, cats, wild animals and also affect human. This research aimed to explore the protein in mites S. scabiei which has antigenic character and play roles in scabies pathogenesis in goats and rabbits. METHODS: S.scabiei mites were isolated from goats and rabbits, and characterized using SDS-PAGE. In addition the protein was also analysed using Western Blot assay. The isolation and identification were carried out in 2015 at the Parasitology Laboratory of Veterinary Medicine Faculty, Universitas Airlangga, Surabaya, Indonesia. RESULTS: The identification results using SDS-PAGE of mites S. scabiei var. caprae expressed 12 protein bands between 26,7 kDa and 205,8 kDa, continued by Western Blot showed 3 protein bands, after being reacted with blood serum from scabies infected goat, it could be identified antigenic protein with molecule weight 205.8 kDa, 57.3 kDa, and 43 kDa. While protein in mites S. scabiei var. cuniculi identified 9 protein bands between 24 kDa and 75 kDa by SDS-PAGE, and the Western Blot assay identified antigenic protein with molecule weight 62 kDa and 51 kDa. CONCLUSION: The antigenic protein of S. scabiei var. caprae and S. scabiei var. cuniculi showed that they are probably involved in the scabies pathogenesis in goats and rabbits.

Morphologic and morphometric characteristics of ascaroid worm, Ophidascaris piscatori in Xenochrophis piscator snake in Sidoarjo, Indonesia.

AIM: The study was conducted to describe the morphology and morphometry of nematode worm in the stomach of Xenochrophis piscator snake macroscopically and microscopically using light and scanning electron microscopy (SEM). MATERIALS AND METHODS: The sample was 40 nematode worms that have been collected from 60 snakes which slaughtered at snake slaughterhouses in Sidoarjo, Indonesia. The worms (10 male and 10 female) were stained with carmine, and the others were sent to Indonesian Science Institute for ultrastructure observation by SEM. Some of the female worms were immersed in physiological NaCl and incubated to collect the worm eggs. RESULTS: Nematode worm in this study had three lips with almost the same size and it had papillae, so it was included in ascaridoid. The mouth of ascaridoids has three lips, the dorsal bearing two large outer papillae and the each lateroventral with one papilla. The body length and width of the male worm were 70-105 mm and 0.92-1.32, respectively, with head diameter of 0.22-0.28 mm. Dorsal and ventrolateral lips almost have the same size that was 0.10-0.12×0.11-0.13 mm. The length of interlabia was 0.06-0.08 mm, esophagus was 3.21-4 mm, tail was 0.17-0.23 mm, and spicule was 2.12-3.36 mm. The body length and width of the female worm were 85-130 mm and 1.28-1.71 mm, respectively. The head diameter was 0.29-0.38 mm. Dorsal and ventrolateral lips almost have the same size that was 0.13-0.16×0.15-0.19 mm. The interlabial length was 0.08-0.10 mm, esophagus was 3.04-4.67 mm, and tail was 0.22-0.31 mm. The distance of the vulva from the anterior edge was 56-88 mm with an average of 67.35 mm. The eggs have conspicuously pitted with length 0.08-0.09 mm and width 0.07-0.08 mm. CONCLUSION: Based on the characteristics of morphology and morphometry, the ascaroid worms found on X. piscator snake from Sidoarjo, Indonesia, were O. piscatori.

Humoral and cellular immune response induced by antigenic protein of Sarcoptes scabiei var. caprae.

AIM: Scabies is one of the most important diseases in goats and caused by a complex hypersensitivity process that involves both humoral and cell-mediated immune responses. This phenomenon shows that the variety of Sarcoptes scabiei has different characteristics of specific antigenic protein or different immune-dominant. This research aims to detect the humoral and cellular immune response of rabbits which were immunized with the protein of S. scabiei var. caprae. MATERIALS AND METHODS: This research was done as follows, identification and collection of Sarcoptes scabiei var caprae from goat infected with scabies, separation of protein antigen from S. scabiei mites with ultrasonic sonicator, measurement of protein content with spectrophotometry, rabbit injection with 500 µg dose of antigen protein which was repeated 5 times (5x booster) every 2 weeks. Measurement of IgG titer using indirect ELISA, whereas to detect the expression of cellular immune response (TLR-9, CD4, and CD8) using Direct Immunofluorescence assay. RESULTS: Based on the statistical analysis, it showed that there was a significant enhancement (p<0.05) for optical density value or antibody titer and cellular immune response was shown by TLR-9, CD4, and CD8 expression in rabbit T lymphocytes which appear yellow to green fluorescent color using fluorescence microscope. The amount of fluorescence T lymphocytes showed a significant difference (p<0.05) between the control and various boosters. CONCLUSION: Antigenic protein of S. scabiei var. caprae contains ligands, which are involved in the pathogen-associated molecular pattern that has an ability to induce humoral and cellular immune response in rabbit. Specifically, that TLR-9 is not only involved in innate immunity but also in adaptive immunity and can be used as alternative adjuvant development research.

INCREASED APOPTOSIS SKULL OF PUPS BORN TO TOXOPLASMA GONDII-INFECTED MICE ASSOCIATED WITH INCREASED EXPRESSION OF INTERFERON GAMMA, BUT NOT TUMOR NECROSIS FACTOR ALFA.

BACGROUND: Toxoplasma gondii is an intracellular obligate protozoan parasite that infects most warm-blooded animals including humans. It can cause congenital infection with clinical symptoms ranging from mild to severe including microcephaly. At the cellular level, infection T. gondii causes apoptosis in some tissues and it is induced by proinflammatory cytokines, such as IFN-γ and TNF-α. The purpose of this study is to determine the role of proinflammatory cytokines (TNF-α and IFN-γ) to apoptosis skull of newborn from T. gondii-infected mice. MATERIALS AND METHODS: Twenty pregnant mice were divided into two groups. The first group was the control group which was not infected with T. gondii tachizoites. The second group was the infected mice, which was infected with T. gondii tachizoites on the day 11.5 of gestation. All mice were cared until delivery. Subsequenly, pups of the mice were sacrificed and their skullcap tissues were taken for histological preparation. The tissues were stained by TUNEL Assay and IHC. Observed variables were apoptotic index and the percentage of skull cell expressing TNF-α and IFN-γ. Data were analyzed with t-test and regression. RESULTS: Compared to the control group, the skull of the pups born to T. gondii-infected mice showed that the number of apoptotic index and percentage of expressing TNF-α and IFN-γ cells were higher than the control group. There is no correlation between increasing expression of TNF-α and apoptosis skull of pups. However, an increasing expression of IFN-γ affected the increased apoptosis of skull pups born to T.gondii-infected mice. CONCLUSION: Congenital toxoplasmosis in mice increased apoptotic index of skull and the apoptosis of skull associated with increasing expression of IFN-γ, but not associated with increasing expression of TNF-α.