Prevalence of Trypanosomiasis in Wild Rats in Banyuwangi District

The aim of this research was to investigate the number of prevalence of Trypanosomiasis of wild rats in Banyuwangi District. Sixty wild rats were trapped from 4 sub-districts, Banyuwangi, Srono, Songgon, and Tegaldlimo in human residence, markets and rice fields from March until June 2017. Blood were taken after anaesthetized using ether. The examination of parasite used blood smear in Giemsa staining. The result show that just 1 of 60 blood sample was appear Trypanosoma sp. , it means the prevalence of Trypanosomiasis of wild rats in Banyuwangi was 1.67%. Rat-Associated


INTRODUCTION
Trypanosomiasis is a disease which has important role in human and veterinary medicine. This disease caused by Trypanosoma sp., known as protozoan parasite which has flagella in their body (Dobigny et al., 2010).
In Thailand, Trypanosomes which have quite similar morphological parameters with T. lewisi-like were found from an infant with fever, cough, also anorexia (Sarataphan et al., 2007). As mentioned by Shegokar et al., (2006) in India T. evansi was found for the first time in human around 2004. These cases possibly the effect of rodents and wild rats' existence in human dwelling. As the population of wild rats has getting bigger nowadays, it mostly lives in close area with human. In some case wild rats become the first competitor of humans for food, mostly about the preharvest damage. This condition is true according to Meerburg (2009), has mentioned in Malaysia, the precentage of crop losses has reached 5%. The magnificent number of crop losses happened in Indonesia, it reached 17%. The huge number of crop losses in both country has shown that rodents existence could be some kind of threat to human, in this case is about food.
In Indonesia, Trypanosomiasis mostly found in livestock, which infecting cattle. Any report about wild rats Trypanosomiasis in Indonesia is almost none until now (Sim and Wiwanitkit, 2015). Study on prevalence of Trypanosomiasis in wild rats caused by T. lewisi has been done in many countries since mid-20 th century, first was conducted in New Zealand, USA, and more infection has been reported from African and Asian countries, South America also have reports in Chile and Brazil (Linardi and Botelho, 2002).
According to Suwanti and Mufasirin (2014), in their research about wild rats Trypanosomiasis through 2011 until 2014, 7 out of 89 wild rats were infected Trypanosoma sp. in Surabaya.
Wild rat has become serious problems which related to public health and rodent-borne diseases. Wild rats also have been acknowledged as a vector of some diseases, mainly zoonotic diseases. Wild rats have known as a host of more than 60 zoonotic diseases that becomes the top threat to human health as mentioned by Blasdell et al. (2015) in Meerburg et al., (2009). Many rodents borne diseases that have ever been recorded in the world according to Nurisa and Ristiyanto (2005). There are 14 diseases that caused by protozoa. One of these 14 rodent borne diseases is Trypanosomiasis, which mostly happened in the tropical area of the world and possibly transmitted to human.
Banyuwangi District is one area that also has potential to conducted Trypanosomiasis detection in wild rats. Because Banyuwangi District is known as one of the endemic area of Trypanosomiasis in accordance with the research of Sawitri et al., (2015), which had been taking the isolate of Trypanosomiasis from some areas in Indonesia to represent the endemic area, including Banyuwangi. It has been decades since Trypanosomiasis or Surra had entered to Banyuwangi District.
Banyuwangi District has tropic condition which is relatable to the occurrence of Trypanosomiasis, mostly happen in tropical area. Viewing the terms of territory and epidemiological conditions, it has the potential to also become home to a Trypanosomiasis that attack wild rats. Based on literacy, many researches have been done about the prevalence of parasites in wild rats around the world. Taiwan, United States of America, until Malaysia, they have been doing this research for quite long time (Shafiyyah et al., 2012).
Therefore, since there has not been many research with this topic in Indonesia, especially in Banyuwangi District and according to the background above, the author has considered this research about Trypansoma sp. infection of wild rats which has zoonotic effect to human needs to be done in Banyuwangi District.

Samples
This research was Cross-Sectional survey. This research used 60 bloods of wild rats as research samples.

Preparation of blood collecting
Blood collecting process were done after the wild rat have anaesthetized. The blood collected through cardiac puncture using 3 ml tuberculin syringe. Then the blood transferred to EDTA tube quickly before clotting.

Preparation of blood smear
To make blood smear through some steps. First step, blood drop to the object glass using pipette, in the one side or nearly at the edge of the first object glass. Another object glass is put in front of the dropped blood until it is wide spread in the second object glass. Positioning the object glasses form a ± 45º. The second object glass is pulled in front until formed a thin layer of blood.

Preparation of blood staining
Blood smear were fixated in methanol absolute 96%, for 3 minutes. Staining started after fixation by using Giemsa stain 10% for 30 minutes in staining jar. Blood smear films are washed by flowing water or sterile aquades and dried.

Microscopic examination
Microscopic observation of peripheral blood smear was used a microscope with 1000 magnification with oil emersion and then captured with OptiLab®.

Data analysis
Data was analyzed with descriptive method based on the formula of prevalence.

RESULTS AND DISCUSSION
In this research were used 60 wild rat blood samples to know the prevalence of Trypanosomiasis in wild rats in Banyuwangi District. Wild rats were taken from human residence, rice fields, garden, or traditional markets, representing four sub-districts in Banyuwangi, such as Banyuwangi, Srono, Songgon, and Tegaldlimo, from March to June 2017. One of 60 blood samples was positive, the number of prevalence was 1.67% as seen in the Table 1 and Figure 1 below.
To make a clearer view of Trypanosoma sp. as captured in Figure  1, the morphology of trypanosome that was found in wild rat's blood was described in a sketch like in Figure 2 below.
Based on the research result, only 1 sample postive from 60 blood samples that had taken from wild rats. The overall prevalence is 1.67%. The result was shown by the representative four sub-districts in Banyuwangi. It is lower from Surabaya, which had 7,9% according to Suwanti and Mufasirin (2014). They detected 89 wild rats in Surabaya started from 2011 until 2014.
In this research, choosen 4 subdistricts shown the prevalence of Trypanosomiasis in wild rats which was 1.67%. That one positive sample was found in human residence area in Kalipait Village, Tegaldlimo Subdistrict. While in the other sub-districts were not found.
The number of prevalence shown that in different area or region, the result will be out differently. It shows that the prevalence of Trypanosomiasis of wild rats is not only affected by the species or individual factors, but also external factors such as vector development, habitat conditions, include temperature and humidity.
Linardi and Botelho (2002) (2002), T. lewisi has long thin posterior end with subterminal ovale kinetoplast, nucleus was located in the anterior part of the body with free flagellum. The trypanosome that was found has large body, it might be present in intermediate form with migration of the kinetoplas and granular body. This result depends on the incubation period.
The trypanosome was only one single trypanosome in the figure. As mentioned by Desquesnes (2013), based on the development condition and the  Shegokar et al., (2006) T. evansi was found for the first time in human. In 2010 also found T. lewisi infection in a 37-days old infant in India (Verma et al., 2011). Truc et al. (2013) also mentioned, that Trypanosomiasis in human has occurred at Malaysia in 1933, Sri Lanka in 1999, and mentioned that most of the records were come from India.
Study about the infection of Trypanosomiasis of wild rats has not been done oftenly in Indonesia and it was the first time to be done in Banyuwangi District. This should become a warning that the infection of Trypanosomiasis in wild rats has a potential to infect human.

CONCLUSION
It could be concluded that the number of prevalence of Trypanosomiasis in Banyuwangi District was 1.67%. T. lewisi predicted as the species of Trypanosoma sp. that was found. Trypanosoma sp. might still potential to cause outbreak, especially in cattle because Banyuwangi District is known as the endemic area of Trypanosomiasis or Surra in cattle. It also possible to infect human since it was found in human residence area.