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*Corresponding author’s e-mail: drmkalex@yahoo.com1Department of Virology, Faculty of Veterinary Medicine, Mehmet Akif Ersoy University, Burdur, Turkey.2Department of Pathology, Faculty of Veterinary Medicine, Mehmet Akif Ersoy University, Burdur, Turkey.

Pestivirus infections in kids of wild goats (Capra hircus aegagrus)S. Hasircioglu1, O. Ozmen2, M. Kale1,* and H.S. Saltik1

Department of Virology, Faculty of Veterinary Medicine,Mehmet Akif Ersoy University, Burdur, Turkey.Received: 09-05-2016 Accepted: 07-11-2016 DOI:10.18805/ijar.v0iOF.7260

ABSTRACTFive stray wild goat kids (1.5-6 months old) found in forestland by Antalya Directorate of Nature Conservation and NationalParks within different periods were brought to Antalya Zoo for feeding and care. The kids were dead after a short time oftheir arrival to the zoo. The dead animals were examined within the same day of their arrival to the lab. During the viralexamination, presence of pestivirus antigen (Ag) was detected in blood samples, brain and liver of months old kid, brainand spleen of two animals viz. six months and two months old. All tissue samples were also studied for Bluetongue virus(BTV) Ag by Fluorescent Antibody Test (FAT) but the samples were found to be negative. While two animals of six and twomonths old showed Pestivirus infection according to their clinical symptoms, virological and pathological results.

Key words: Antigen ELISA, Bluetongue virus, Pestivirus, Wild goat.

INTRODUCTIONPestiviruses have been detected in more than 50

species of wild animals living free in natural life in variousparts of the world (Campen et al. 2001; Arnal et al. 2004;Vilcek and Nettleton, 2006; EAZA, 2014; Ridpath, 2015).Infection during the first trimester can produce calves thatremain persistently viraemic for life (Brownlie, 1990).Persistent animals also show such immunotolerance againstinfectious virus strain that although they look healthy, theanimals might infect other animals spreading the virus withall their secretions and excretions (Lindberg and Houe,2005). Even if the reservoir state of pestiviruses in wildanimals is not completely clear, studies carried out in thisdirection have drawn attention recently (Casaubon et al.2012).

Bluetongue virus (BTV) infection is important viraldisease transmitted by stinger flies and cause seriouseconomic losses especially in small ruminants (Murphy etal. 1999; Roy, 2010). Infectious particle of BTV firstlyspreads into lymph nodes and moves towards epithelialtissues with affinity, then spreads onto spleen, liver, othertissues and organs. During the last stage of the infection, thevirus runs in blood and might stay here also (Roy, 2010).During 28-56 days of pregnancy in small ruminants, Akabanevirus (AKAV) carries out its replication first in placentaendothelial cells, then in trophoblastic cells and finally infetus itself and causes congenital defects, arthogryposis (AG)hydrancephaly (HE) and abortions (Murphy et al. 1999;Mellor and Kirkland, 2008). Even though the host spectrumhasn’t been revealed completely, Peste des Petits Ruminants

virus (PPRV), known to have caused acute, high mortalityand contagious diseases in pure sheep and goat populations,doesn’t provide sufficient information about its pathogenesisfor wildlife and zoo animals (Murphy et al. 1999; Barrett,2008). Specific antibodies against BTV and AKAV havebeen detected in many animals living in wildlife (such ashorses, pigs, deer, antelopes, hippos, giraffes etc.), and widewildlife host spectrum is suspected for PPRV (Barrett, 2008).

In this study, after the death of wild mountain goatkids found alone in wild and taken to Antalya Zoo to becared, presence of Pestivirus and BTV was studied.MATERIALS AND METHODSClinical and/or necropsy samples: Five strayed mountaingoat kids as one six-months-old female (No:1), one 1.5-months-old male (No:2), one 1.5-months-old female (No:3),one two-months-old female (No:4) and one two-months-oldfemale (No:5) found in forestland by Antalya Directorate ofNature Conservation and National Parks within differentperiods were taken to Antalya Zoo for feeding and care. Allthese animals died in 1-2 days due to diarrhea, weaknessand anorexia. The dead animals were presented to pathologyand virology labs of our faculty with cold chain within thesame day. The animals were sent in the spring of 2015.

Blood was collected from the hearts of kids havingdied in pathology necropsy hall. Serum and leucocytes werethen separated from the blood samples. Samples of brain,liver, spleen and lung from all animals were collected intosterile plastic containers by using different sterile clippersand bistoury for each animal.

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Necropsy was performed for all five animals. Tissuesamples were collected and fixed in 10% buffered formalinfor histopathological examination. Then tissue samples wereroutinely processed and blocked in paraffin. Five micronwas sectioned by a Leica 2155 rotary microtome and stainedwith hematoxyline-eosin (HE) and examined under the lightmicroscope (OIE Manuel of Diagnostic Tests and Vaccinesfor Terresrial Animals 2012).Antigen ELISA for Pestiviruses: Bovine Viral DiarrheaVirus (BVDV) ELISA (IDEXX, USA) kit was used to detectpestivirus antigen (Ag) presence in leucocyte samples of allanimals.Fluorescent antibody tests (FAT) for Pestiviruses andBTV: Cross sections of tissue samples (brain, liver, spleen,lung) were palpated separately onto each preparate for thestudy. Having been dried for at least 30 minutes in roomtemperature, the preparates were fixed in acetone for at least15 minutes again in room temperature. After evaporatingthe acetone on preparates completely, spesific 50-75 µl Anti-FITC-Ig (anti-fluorescein isothiocyanate-immunglobulin)conjugate was performed on preparates for each factor.Having been left for incubation for 30 minutes at 37 °C,the preparates were then kept in Rinse Buffer, pH 9.0(VMRD catalog no 210-90-RB). After drying completely,one single drop of 90% glycerin 10% PBS (pH 7.4)mixture was dropped onto preparates and the area to bescanned by the help of dropper and this area was closed

with a coverglass. All preparates were studied underfluorescent microscope.RESULTS AND DISCUSSION

At the end of the virological test performed forleucocyte samples, pestivirus Ag (+) for sample No:1 and 4,and Ag(-) for others was detected (Table 1).

At the end of FAT performed for tissue samples,pestivirus Ag (+) in brain and liver tissues and pestivirus Ag(-) in spleen and lung tissues for sample No:1 was found. Inall tissues of this sample, BTV Ag (-) was found (Table 1).Pestivirus Ag (+) in brain and spleen tissues and pestivirusAg (-) in liver and lung tissues for sample No:4 was detected.In all tissues of this sample, BTV Ag (-) was found(Table 1).

At necropsy, slight hyperemia was observed inmesenterial vessels in all animals. In two kids (No:2 and No:3)approximately 20-30 ml serous, pink color fluid was seen inabdominal cavity. Watery content was observed in the lumenof the intestine. Marked hyperemia and slight hemorrhagicfoci was noticed in meninges in two animals (No:1 andNo:4). There was no congenital abnormality in any organ.

At the histopathological examination, the mostcommon lesions were localized in brains. In addition tomeningeal hyperemia, edema, hemorrhages and slightlymphocyte infiltrations were observed around the vessel ofbrains in two kids (No:1 and No:4) (Figure 1). Hyperemia

Samples ELISA (Pestivirus-Ag) FAT (Pestivirus-Ag / BTV-Ag)

Brain Liver Spleen Lung

No:1 + + / - + / - - / - - / -No:2 - None None None NoneNo:3 - None None None NoneNo:4 + + / - - / - + / - - / -No:5 - None None None None

Table 1: The results of Ag ELISA and FAT

Fig 1: Histopathology of brains of kids that died from pestivirus infection (A) Slight perivascular lymphocyte infiltration (arrows),(B) Slight perivascular infiltration (arrows) and hemorrhage (arrow heads), HE, Bars=100µm.

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and degenerations, slight lymphocyte infiltrations andKupffer cell proliferations were observed in one kid’s liver(No:1). In all kids, increase in septal wall thickness and slightinflammation was found in lungs (Figure 2). Desquamationand slight infiltration was noticed at the intestinal propriamucosa. There were no pathological findings in other organsexcept hyperemia.

The permanent changes that humans make on earthin line with socioeconomy, together with animaldomestication, global warming and natural events mighttrigger the interactions (describe the kind of possibleinteractions) between wild and domestic animal species.Even though these interaction mechanisms couldn’t beclarified completely in terms of viral infections, in somestudies, these kinds of interactions were reported to haveplayed an important role on the pathogenesis of infections(Bengis et al. 2002).

In a study performed by Pastoret et al. (1988) onpestivirus infections in wild ruminants, they reported thatthere was no connection of the pestivirus infection betweenwild animals and domestic ruminants. After that, Meyling etal. (1990) stated that the main resource of pestivirusinfections in domestic ruminants is wild animals. Hence,Becher et al. (1997) determined that pestiviruses have a fairlycommon host spectrum in biungulates (Artiodactyla) whichthey could infect passing through the barrier between speciesand that especially BVDV and BVDV could infect manydomestic and wild ruminants (Becher et al. 1997; Becher etal. 2003). Mishra et al. (2007) reported the presence ofpestivirus in Native Indian goats (Capra hircus) and evenisolated BVDV type 2 strain.

Over the past years in our country BTV infectionpresence was detected in Western Anatolia region and wasbrought under control by a successful vaccination programme(Yonguc et al. 1982). Besides, during a serological studyperformed in Black Sea region, antibody (Ab) presence in

domestic ruminants was found (Albayrak and Ozan, 2010).The fact that wild ruminants also were in the host spectrumof BTV is one of the realities of this study. BTV antibodypositive results determined in this study (No:1-5 bloodsamples) points to the presence of BTV infection in wildlife.However, whether there is a connection between BTVinfections found in domesticated goats and wild mountaingoats still isn’t known corporally yet.

Domestication of Capra hircus aegagrus, knownas the ancestor of domestic goats around the world, for thefirst time in Turkey, Northern Syria, Mount Zagros, Iran andIraq regions in 10000 BC (Zeder and Hesse, 2000) showsthat the infections lately detected in these regions might infact date back to earlier years. Accordingly, viral infectionsshowing similar symptoms and each condition andenvironment (mutual grazing lands, vectors etc.) where wildand domestic goats could contact each other would play animportant role in interrelated infection. Together with theobtained results, the fact that the clinical symptoms found infree-range mountain goats in their late stages and thepathological findings from internal organ samples ofnecropsy animals showed a similarity with the table ofinfection that viral diseases caused in domestic cattle, sheepand goats was such as to support Pestivirus (Ag) and BTV(Ag) findings detected in this study.

At the end of this study, we realize that we need tostruggle in wildlife also in order to prevent Pestivirus andBTV infections. For this purpose, we need to isolate waterand grazing areas of domestic sheep-goat populations andestablish buffer zones to prevent domestic animal populationsfrom infections through possible contact routes. Besides, forthese diseases, we suggest to establish combat programmesin wildlife similar to the one for rabies.ACKNOWLEDGEMENT

We thank Aygül ARSUN, Dr. (Antalya Zoo) andother staff for assistance with animals.

Fig 2 (A): Slight lymphocyte infiltration (arrows) and increase in Kupffer cels (arrow heads) in a liver a kid died from pestivirusinfection, (B) Hyperemia, increase in septal wall thickness (arrow heads) and slight mononuclear cell (arrows) infiltrations in lungs,HE, Bars=100µm.

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