may-jr (artigo) hematology and blood chemistry parameters differ in free-ranging maned wolves

7/28/2019 May-jr (Artigo) Hematology and Blood Chemistry Parameters Differ in Free-ranging Maned Wolves

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HEMATOLOGY AND BLOOD CHEMISTRY PARAMETERS DIFFER IN

FREE-RANGING MANED WOLVES (CHRYSOCYON BRACHYURUS)

LIVING IN THE SERRA DA CANASTRA NATIONAL PARK VERSUS

ADJACENT FARMLANDS, BRAZIL

J. A. May-Junior,1,5 N. Songsasen,2,5 F. C. Azevedo,1 J. P. Santos,1 R. C. Paula,1,3 F. H. G.Rodrigues,1,4 M. D. Rodden,2 D. E. Wildt,2 and R. G. Morato3,6

1 Projeto Lobo-Guara, Instituto Pro-Carnvoros, Sao Paulo, Brazil2 Conservation and Research Center, Smithsonians National Zoological Park, Front Royal, Virginia, USA3 Centro Nacional de Pesquisa para a Conservacao dos Predadores Naturais-CENAP-ICMBio, Sao Paulo, Brazil4 Departamento de Biologia Geral, Universidade Federal de Minas Gerais, Minas Gerais, Brazil5 These authors contributed equally to this work6 Corresponding author ([email protected])

ABSTRACT: There has been growing interest in the specific impacts of anthropogenic factors onthe health of wildlife. This study examined hematology and serum chemistry status of a prominent

carnivore, the maned wolf (Chrysocyon brachyurus), living in, on the boundaries to, or on adjacentfarmlands to the Serra da Canastra National Park, Brazil. Twenty-eighty wolves were captured,and values were compared 1) between subadults (n58 animals) and adults (n520 animals), 2)males (n512 animals) and females (n516 animals), and 3) among wolves living inside the park(n511), near the park border (n511 animals), and in neighboring farming areas (n56 animals).Age, gender, and wolf locations influenced (P,0.05) hematology and serum biochemistry values.Specifically, adults had lower (P,0.05) circulating phosphorus than subadults. Males had lower(P,0.05) serum glucose, creatinine phosphokinase, and cholesterol and higher (P,0.05)potassium than females. Erythrocyte count and serum cholinesterase were lower (P,0.05) inwolves living within the park compared with near the park border or on farmlands. Meancorpuscular volume was lower (P,0.05) in wolves living near the park border than those rangingwithin the park and on farmlands. Aspartate transaminase and chloride were higher (P,0.05) inwolves living inside the park compared with those ranging near the park border. Creatininephosphokinase was lower (P,0.05) in wolves living on farmland compared with the other twolocations. These results clearly reveal a relationship between age and gender on hematology andserum biochemistry values in free-living maned wolves. More importantly, certain traits indicativeof health are potentially compromised in wolves living in areas under anthropogenic pressure.These data lay a foundation for examining the influence of farming and local domestic species ondisease susceptibility and fitness in the maned wolf.

Key words: Blood biochemistry, free-ranging, health, hematology, maned wolf.

INTRODUCTION

The maned wolf (Chrysocyon bra-

chyurus) is the largest canid (2030 kg)of the Cerrado (grassland) ecosystem inthe Brazilian Central Highlands (Dietz,1984). The species also ranges in Argen-tina, Paraguay, Bolivia, and Peru (Roddenet al., 2004), but its population status is not

well defined. Although only listed as nearthreatened by the International Unionfor Conservation of Nature (IUCN)-

World Conservation Union (2006), the

wild population of maned wolf is declin-ing, probably due to conversions of habitatto farmland. During the last 30 yr, morethan 70% of the Cerrado have been

radically modified for agricultural purpos-es (Myers et al., 2000).

Over the last decade, there has been a

growing interest in the relationship be-tween human-induced alterations in hab-itat availability/quality and the health of

wildlife living in or adjacent to suchlandscapes (Laurenson et al., 1997; Das-zak et al., 2000; Cleaveland et al., 2001;Gillin et al., 2002; Lanfranchi et al., 2003).Such encroachments increase the likeli-hood of direct or indirect contact betweenhumans and their domestic animals and

wildlife that, in turn, increases risk ofdisease transmission across all species(Lanfranchi et al., 2003). There now areclassical examples for the legitimacy of

Journal of Wildlife Diseases, 45(1), 2009, pp. 8190# Wildlife Disease Association 2009

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such concerns, for example, the transmis-sion of the rabies pathogen from domesticdogs to the Ethiopian wolf (Canis simen-

sis) and African wild dog (Lycaon pictus),both of which resulted in significant

population declines (Sillero-Zubiri andMacdonald, 1996; Woodroffe et al.,1997). To date, similar studies have notbeen conducted for any Latin Americancarnivore. Although there is no directevidence of disease in Brazilian free-ranging maned wolf populations, a recentstudy in Bolivia has shown that wildindividuals of this species have beenexposed to common infectious diseases of

domestic dogs (Canis familiaris; Deemand Emmons, 2005). The pathogensinclude canine distemper virus, canineparvovirus, rabies virus, canine coronavi-rus, canine adenovirus, Leptospira inter-rogans, Toxoplasma gondii, and Dirofilar-

ia immitis. Therefore, the maned wolf inother regions, including Brazil, may wellbe at risk for multiple diseases.

A prerequisite to examining the sensitiv-ity of any wildlife species to disease is firstcharacterizing baseline normal values andgeneral health status (Karesh et al., 1995).To date, health-related data have beengenerated for wild maned wolves, includinginformation on hematology, blood chemis-try, and incidence of parasitic infestations(Dietz, 1984; Mattos, 2003; Deem andEmmons, 2005). No systematic study hasbeen conducted to explore the influence ofage, gender, and adjacency to human

development/activities on hematology andblood chemistry normal values. Interest-ingly, there is a substantial database onhealth issues associated with maned wolvesliving ex situ in zoological collections thatspans almost 30 yr (Fletcher et al., 1979;Bush, 1980; Norton, 1990; Maia andGouveia, 2002). This includes detailedinformation on species hematology andserum biochemistry (Barbiers and Bush,

1994) and overall sensitivity to rabies,canine distemper, and parvovirus (Fletcheret al., 1979; Norton, 1990).

Our ultimate aim is to understand

whether rapidly encroaching farmingpractices and land conversions are in-creasing disease risks in maned wolves inthe Cerrado ecosystem. Thus, it is neces-sary to establish hematology and blood

chemistry values for males versus females,adults versus subadults, and wolves thatlived exclusively in a protected area versusthose range on park borders or surround-ing farmlands. Our central target area ofstudy was the Serra da Canastra NationalPark that is rich in biodiversity, but alsounder significant development pressure,largely for cattle ranching and coffeeplantations. We considered wolves sam-

pled within the park as being minimallydisturbed by humans. In contrast, wolvescaptured near the park boundary and onneighboring farmlands were considered tobe under marginal or maximal disturbanceand/or stress, respectively.

MATERIALS AND METHODS

Study area

The study was conducted within the Serra

da Canastra National Park and surroundingareas (46u159W, 20u009S) in the Minas Geraisstate of southeastern Brazil. The park is aprotected area encompassing 2,000 km2, with715.2 km2 regulated and managed by theInstituto Chico Mendes de Conservaao daBiodiversidade (ICMBio), and the remainderwas privately owned. This largely is an area ofopen grasslands with variety of vegetation,including some spots with Cerrado (savannah),small bush around the creeks, stone fields, andsmall areas of rain forest.

Animal capture, anesthesia, andphysical evaluation

Twenty-eight maned wolves (12 males, 16females) were captured from January 2004through December 2006 using box trapsbaited with chicken or fish. These traps wereplaced at locations within the park, near thepark border (within 5 km of the park border),and on privately owned farms (from 5 km to30 km from the park border). Each trap waschecked daily. Once captured, each animalwas chemically restrained using tiletamine-zolazepan (4 mg/kg; TelazolH, Fort Dodge,Sao Paulo, Brazil; Bush, 1996; Kreeger, 1999)administered by hand syringe. Vital signs,including body temperature, heart rate, and

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respiratory rate, were recorded every 15 minthroughout the 60-min anesthesia. Each wolfwas weighed using a portable scale (Pesola,Baar, Switzerland). Wolf age was estimated byphysical characters (e.g., weight, scars, coatappearance, and dentition) according to the

criteria of Dietz (1984). As a result, eightanimals (five males, three females) were cate-gorized as ,1 yr of age (i.e., subadults), and theremaining individuals (7 males, 13 females)were categorized as.1 yr of age (adults). Basedon overall appearance, each wolf was given anumerical condition score (13, largely based onfat covering, with a 3 rating for wolves with thebest conformation and slightly detectable fatcompared with a 1 rating that was indicative ofpoor condition and no fat covering). Wolveswere also examined for tick (Amblyomma sp.)

infestation and were subjectively classifiedbased on numbers of ticks or fleas present ashaving high, medium, low, or no infestation.Blood and urine samples were obtained (asdescribed below), and 27 of 28 animals werefitted with a radiocollar (Advance TelemetrySystem Inc., Isanti, Minnesota, USA; TelonicsInc., Mesa, Arizona, USA; Wildlife MaterialsInc., Murphysboro, Illinois, USA) for parallelassessments of home range. Subsequent radio-telemetry also allowed confirming the appro-priate categorizing of wolf locations (if wolvesexclusively or predominantly lived within thepark, in the area of the parks boundary, or onfarmlands; see below). After completing theexamination and collecting all samples, eachwolf was monitored to ensure uneventfulrecovery within the box trap from anesthesia.When an animal seemed to be recovered(usually 4 to 5 hr postinjection), each wolf wasreleased from the box trap into its original areaof capture. This research was conducted withinlegal requirements (licenses 108/2006, DIREC,IBAMA) and under license 147/05 and 356/06,CNPq (N.S., M.D.R., and D.E.W.).

Blood sample collection and analysis

For each individual, a 38-ml sample of bloodwas collected from the sephanous vein using aVacutainerH needle (BD Biosciences, FranklinLakes, New Jersey, USA) attached with anappropriate collection tube. Twenty-eight bloodsamples were available for assessment. For eachanimal, 12 ml of the sample was collected intotubes containing EDTA (Sistema Vaccum II,Vacuum II, Villfend Corporation Industria ecomercio Ltda, Itupeva Brazil), 4 ml of whichwas used for hematology analysis in this study,and the remaining sample was stored for otherstudies. Twenty-eight milliliters of blood sam-ples was collected into a tube voided of an

anticoagulant to obtain serum; 4 ml of serumwas used for biochemistry analysis, and the restwas stored for future serologic studies. All bloodsamples were stored in cool boxes (48 C) forthe 30- to 90-min transport to the research base.Aliquots with no EDTA were centrifuged at

5,0003G for 7 min (Q222-T centrifuge, Qui-mis, Sao Paulo, Brazil), serum was recovered,placed in labeled cryotubes, and maintained at220 C until analysis. Blood and sera weretransported (23 hr by car) in cool box (48 C)to a commercial laboratory (Instituto de Pato-logia Clnica Carlos Chagas, Araxa, MinasGerais, Brazil) for automated and consistentassessments of hematology (KX21N, Sysmex,Kobe, Japan) and serum biochemistry (ExpressPlusH, Chiron Corporation, Charlotte, NorthCarolina, USA).

Confirmation on animal location

Our parallel observations of home range anddistribution via radiotelemetry allowed confirm-ing our ability to assign a location category toeach wolf. Originally, we captured 11 wolveswithin the park, 11 living near the park border,and six on farmlands. Each wolf was tracked byradiotelemetry for duration of 1 mo to 2 yr.Subsequent home range measurements con-firmed that these wolves almost always retainedsite fidelity, remaining in their original location

areas. Occasionally, wolves on farmlands strayedfor short intervals (1 day to 2 wk) near the park,but then returned to farms. Wolves within thepark always remained in the protected area.

Statistical analysis

Data are presented as means6standarderror of mean (SEM). Statistical analyses wereperformed using SigmaStat software (SPSSInc., Chicago, Illinois, USA). Comparison ofmean hematology and blood biochemistryvalues on the basis of gender and age were

performed using a Students t-test. Compari-son of mean hematology and serum biochem-istry values among individuals living within thepark, near the park border, and on farmlandswas performed using one-way analysis ofvariance (when data were normally distribut-ed) or Mann-Whitney U-test (for non-normaldistributed data). Differences among groupswere determined using Duncans multiplerange test or Dunns test. Differences wereconsidered significant at P,0.05.

RESULTS

All wolves were in good body conditionand seemed overtly healthy. The average

MAY-JUNIOR ET AL.HEALTH EVALUATION OF FREE-RANGING MANED WOLF 83


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body weight of subadults was 22.763.1 kg,which was less (P,0.05) than 26.764.3 foradults (P,0.05). There were no massdifferences (P.0.05) within age groupsbetween the genders (subadults, 23.062.3 kg for males vs. 22.364.6 kg forfemales; adults, 29.161.8 kg for males vs.25.364.7 kg for females).

Hematology values were not affected byage and genders (P.0.05) (Tables 1 and2). Regardless of gender and location, theonly serum biochemistry value influencedby age was increased (P,0.05) phospho-rus concentration in younger compared

with older animals (Table 3). When ageand location were not factored, thengender had a modest impact on serumbiochemistry; females had higher

(P,0.05) glucose, creatinine phosphoki-nase, and cholesterol concentrations butlower (P,0.05) potassium than males(Table 4).

When data were analyzed on the basisof animal location, there was an effect(P,0.05) on erythrocyte count, meancorpuscular volume, aspartate transami-nase, creatinine phosphokinase, chloride,and cholinesterase (Tables 5 and 6). Spe-cifically, erythrocyte count was lower(P,0.05) in wolves within the park thannear the park border and farmlands. Meancorpuscular volume was lower (P,0.05) in

wolves near the park border comparedwith wolves within the park or on farm-lands. Microcytic red blood cells werefound in 10 individuals, seven of which

TABLE 1. Hematology values (mean6SEM and range) in subadult (n58) versus adult (n520) maned wolves(Chrysocyon brachyurus) living within and near the Serra da Canastra National Park (Brazil).

Hematology trait Subadult Adult

Erythrocytes (106/mm3) 4.760.1 (4.25.3) 5.060.1 (4.15.9)

Hemoglobin (g/dl) 12.260.4 (10.813.6) 13.160.3 (10.715.4)Hematocrit (%) 38.761.0 (3443) 40.760.9 (3448)Mean corpuscular volume (fl) 82.161.4 (7688) 80.860.8 (7689)Mean corpuscular hemoglobin (pg/cell) 26.160.5 (2327) 26.060.2 (2428)Mean corpuscular hemoglobin concentration (g/l) 31.660.5 (3034) 32.160.2 (3034)Leucocytes (103/mm3) 12.360.9 (9.216.8) 12.160.7 (7.919.1)Neutrophils (103/mm3) 7.361.1 (3.512.6) 8.660.6 (5.516.1)Eosinophils (103/mm3) 0.960.1 (0.41.4) 0.760.1 (01.7)Basophils (103/mm3) 0.0160.01 (092) 0.060.0 (0)Lymphocytes (103/mm3) 3.460.9 (1.89.6) 2.260.1 (0.83.7)Monocytes (103/mm3) 0.660.02 (0.31.3) 0.560.0 (0.11.3)

TABLE 2. Hematology values (mean6SEM and range) for male (n512) versus female (n516) maned wolves

(Chrysocyon brachyurus) living within or near Serra da Canastra National Park (Brazil).

Hematology trait Male Female

Erythrocytes (106/mm3) 5.060.2 (4.15.7) 4.960.1 (4.15.9)Hemoglobin (dl) 13.060.4 (10.814.4) 12.860.3 (10.715.4)Hematocrit (%) 40.661.1 (3644) 39.861.0 (3348)Mean corpuscular volume (fl) 81.061.4 (7689) 81.360.7 (7686)Mean corpuscular hemoglobin (pg/cell) 25.860.4 (2328) 26.160.3 (2428)Mean corpuscular hemoglobin concentration (g/l) 31.960.4 (3034) 32.060.3 (3034)Leucocytes (103/mm3) 11.860.9 (9.219.1) 12.460.7 (8.619.4)Neutrophils (103/mm3) 8.060.8 (5.112.9) 8.260.7 (3.516.1)

Eosinophils (10

3

/mm

3

) 0.76

0.1 (00.9) 0.86

0.1 (01.2)Basophils (103/mm3) 0.00760.007 (092) 0.060.0 (0)Lymphocytes (103/mm3) 2.460.5 (1.53.8 2.761.0 (0.89.6)Monocytes (103/mm3) 0.660.2 (0.31.3) 0.560.1 (0.11.1)



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lived on the park border with the remain-ing individuals being on farmlands. Sub-

jective evaluation of tick infestation foundthat two, seven, and two wolves livinginside the park had medium, low, and noinfestation, respectively. All wolves livingin the park border area were infested withticks, with three, four, and four individualshaving high, medium, and low infestations,respectively. For animals living on farms,five wolves had low tick infestations, withthe remaining wolves having none of theseparasites. Aspartate transaminase, chlo-ride, and creatinine phosphokinase werehigher (P,0.05) in wolves within the parkcompared with those in the border area(Table 6). Among the three groups, cho-linesterase was highest in wolves living onfarmlands and lowest in individuals livinginside the park (Table 6).

DISCUSSION

This is the first systematic evaluation ofthe hematology and serum biochemistry ofthe maned wolf in the Serra da Canastra

National Park, Brazil, as well as in theadjacent habitat areas. These findingsprovide important baseline data for anongoing effort to examine the influence ofrapid human encroachment on diseasesusceptibility and long-term fitness of thisflagship species. Our results are significantbecause of sample size and due to ourexpansive sampling geographyoverthree distinctive areas (within a protectedarea, near the park boundary, and onadjacent farmlands). In addition to adding

a substantial amount of information to thespecies database, we discovered thatselected serum biochemistry values areconsistently influenced by wolf gender.Therefore, age and gender need to beconsidered in interpreting data in thecontext of animal health status. Althoughall wild maned wolves evaluated in thisstudy seemed healthy, it was noteworthythat there were significant variations in

blood traits among wolves on the basis oflocation. Although it is premature toindicate that human perturbations aredirectly responsible for these variations,

TABLE 3. Serum biochemistry values (mean6SEM and range) for subadult (n58) versus adult (n520)maned wolves (Chrysocyon brachyurus) living within and near the Serra da Canastra National Park (Brazil).

Serum biochemistry trait Subadult Adult

Alanine transaminase (IU/l) 47.268.7 (2495) 76.1614.5 (31278)

Aspartate transaminase (IU/l) 39.663.9 (2258) 55.465.3 (14117)Gamma glutamyl transferase (IU/l) 3.360.8 (16) 3.060.5 (05)Glucose (mg/dl) 98.3611.9 (42150) 106.465.0 (71143)Urea (mg/dl) 59.5612.6 (29142) 61.363.6 (3199)Creatinine (mg/dl) 1.060.1 (0.71.4) 1.160.0 (0.71.4)Total protein (g/dl) 7.660.3 (6.38.8) 7.460.3 (4.09.3)Albumin (g/dl) 2.660.1 (2.43.0) 2.560.1 (1.53.5)Globulin (g/dl) 5.060.3 (3.66.2) 4.960.2 (2.56.6)Total bilirubin (mg/dl) 0.360.0 (0.20.3) 0.360.0 (0.10.5)Creatine phosphokinase (IU/l) 178.1647.6 (79442) 297.4636.9 (97624)Lactate dehydrogenase (IU/l) 150.1632.0 (55339) 226.7628.9 (76544)Triglycerides (mg/dl) 34.169.7 (875) 28.265.4 (4299)

Cholesterol (mmol/l) 4.26

0.4 (2.76.01) 4.66

0.2 (3.217.77)Uric acid (mg/dl) 0.4560.2 (0.11.3) 0.4660.1 (0.169)Sodium (mmol/l) 143.761.8 (135150) 144.861.3 (136152)Potassium (mmol/l) 4.560.2 (4.05.3) 4.660.1 (4.05.8)Chloride (mmol/l) 107.862.9 (99117) 106.561.6 (98119)Calcium (mmol/l) 2.460.1 (2.02.7) 2.360.1 (1.83.0)Phosphorus (mg/dl) 6.660.5a (4.28.9) 5.160.3b (3.58.2)Cholinesterase (U/l) 3,095.36294.7 (2,1864,605) 2,976.06232.9 (8595,109)

a,b Different letters within the same row indicate a difference between groups ( P,0.05).



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such findings indicate a need for vigilance

and more detailed study. Indeed, wesuspect that such surveys can serve assentinel information for predicting thepotential of more serious compromises inphysiologic functions, including diseasesusceptibility.

Although maned wolf hematology and

serum biochemistry have been evaluatedextensively in captive populations in zoos(Barbies and Bush, 1994; Maned WolfSpecies Survival Plan, 2007), only limiteddata are available for free-living popula-tions. Dietz (1984) in his original classic

TABLE 4. Serum biochemistry values (mean6SEM and range) for male (n512) versus female (n516)maned wolves (Chrysocyon brachyurus) living within and near the Serra da Canastra National Park (Brazil).

Serum biochemistry trait Male Female

Alanine transaminase (IU/l) 70.1612.1 (24278) 66.1620.1 (3228)

Aspartate transaminase (IU/l) 48.266.1 (2294) 52.965.7 (14117)Gamma glutamyl transferase (IU/l) 2.860.6 (14) 3.360.5 (06)Glucose (mg/dl) 89.566.6a (42119) 11565.6b (71150)Urea (mg/dl) 65.468.5 (29142) 57.364.0 (3190)Creatinine (mg/dl) 1.160.0 (0.81.4) 1.060.0 (0.71.3)Total protein (g/dl) 7.760.3 (6.39.3) 7.360.3 (48.8)Albumin (g/dl) 2.560.1 (22.90 2.560.1 (2.53.5)Globulin (g/dl) 5.260.3 (3.66.6) 4.760.2 (2.56.5)Total bilirubin (mg/dl) 0.360.0 (0.10.4) 0.360.0 (0.10.5)Creatine phosphokinase (IU/l) 181.2641.9a (79542) 324.8638.4b (131624)Lactate dehydrogenase (IU/l) 200.4628.0 (79378) 208.2635.6 (55544)Triglycerides (mg/dl) 38.2610.0 (4118) 23.763.2 (6299)


0.2

a

(2.75.0) 4.86

0.2

b

(3.647.8)Uric acid (mg/dl) 0.560.1 (0.269) 0.460.1 (0.11.2)Sodium (mmol/l) 143.561.1 (135149) 145.261.6 (126152)Potassium (mmol/l) 4.960.2a (4.05.8) 4.460.1b (4.04.8)Chloride (mmol/l) 104.562.0 (96117) 108.761.8 (98119)Calcium (mmol/l) 2.360.1 (1.82.7) 2.260.1 (1.83)Phosphorus (mg/dl) 5.760.4 (3.98.20) 5.560.4 (3.58.9)Cholinesterase (U/l) 2,950.86226.0 (1,9324,705) 3,054.56279.3 (8595,109)

a,b Different letters within the same row indicate a difference between groups ( P,0.05).

TABLE 5. Hematology values (mean6SEM and range) for maned wolves (Chrysocyon brachyurus) livingwithin (n511) versus near the border (n511) versus adjacent farmlands (n56) to the Serra da CanastraNational Park (Brazil).

Hematology trait Within park Near park border Farmland

Erythrocytes (106/mm3) 4.560.4 a (4.15.4) 5.160.1b (4.35.7) 5.360.1b (4.95.9)Hemoglobin (g/dl) 12.060.3 (10.814.6) 13.060.4 (10.714.4) 14.160.4 (1315.4)Hematocrit (%) 38.061.0 (345) 40.761.0 (3444) 43.161.5 (3848)Mean corpuscular volume (fl) 83.860.8a (8089) 79.060.9b (7685) 80.361.5a (7686)Mean corpuscular hemoglobin (pg/cell) 26.560.2 (2628) 25.460.4 (2327) 26.361.4 (2428)Mean corpuscular hemoglobin

concentration (g/l)31.760.3 (3033) 32.060.4 (3034) 32.560.3 (3234)

Leucocytes (103/mm3) 11.860.9 (7.917.7) 11.960.8 (9.119.1) 13.361.5 (8.619.4)Neutrophils (103/mm3) 8.161.0 (5.516.1) 8.060.6 (5.112.9) 8.461.3 (3.512.9)Eosinophils (103/mm3) 0.860.1 (0.01.4) 0.860.1 (0.31.5) 0.760.2 (0.11.7)

Basophils (10

3

/mm

3

) 0.06

0.0 (0) 0.06

0.0 (092) 0.06

0.0 (0)Lymphocytes (103/mm3) 2.260.2 (0.83.8) 2.560.1 (1.83.7) 3.461.2 (0.89.6)Monocytes (103/mm3) 0.560.1 (0.31.3) 0.660.1 (0.31.3) 0.660.1 (0.11.1)

a,b Different letters within the same row indicate a difference among groups ( P,0.05).



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studies of this species in the Serra daCanastra National Park, measured hema-tologic variables from eight wild individu-als (five males and three females). Morerecently, Mattos (2003) measured 26blood traits in 17 wild maned wolves livingin Sao Paulo state, Brazil. Interestingly, allmetrics reported for wild maned wolves(including from the present study) fall

within a general and similar range to

conspecifics held in zoos (Barbiers andBush, 1994).

There were some differences in ourmeasurements on the basis of age as wellas gender. The elevated phosphorus con-centration in subadults has been indicativeof higher bone metabolism that normallyoccurs in younger compared with olderindividuals (Bush et al., 1981; Coles, 1984;Miller and Gonalves, 1996). Male and

female wolves also differed in circulatingglucose and creatinine phosphokinaseconcentrations, which probably reflecteda higher metabolism for females, a phe-

nomenon that occurs at least in thedomestic dog (Heffron et al., 1976).However, unlike the domestic dog (Braunet al., 2003), male maned wolves hadhigher serum potassium than femalecounterparts, demonstrating species vari-ations in certain serum biochemistry traits.

Wolves living near the park borderseemed to have higher levels of tickinfestation than those inside the park and

on farmlands. This may explain theprevalence of wolves on the border alsoexperiencing more microcytic red bloodcells (seven of 10 captured wolves) and alow mean corpuscular volume (MCV). It is

worth noting that the border area hasmore of cattle ranches than coffee planta-tions, the latter making up more of thetrue farmland as categorized in thisstudy. This also could explain the higher

tick infestation in the border area becausethere were more domesticated animalsliving here compared with the coffeeplantations. We suspect that the overall

TABLE 6. Blood chemistry values (mean6SEM and range) for maned wolves living within (n511) versusnear the border (n511) versus adjacent farmlands (n56) to the Serra da Canastra National Park (Brazil).

Serum biochemistry values Within park Near park border Farmland

Alanine transaminase (IU/l) 80.9616.2 (36228) 64.8622.2 (24278) 49.767.9 (3379)

Aspartate transaminase (IU/l) 58.067.7a (31117) 43.966.7b (2292) 50.563.4ab (3960)Gamma Glutamyl transferase (IU/l) 2.560.4 (04) 2.860.7 (15) 4.761.0 (16)Glucose (mg/dl) 111.966.7 (74143) 93.867.5 (42125) 108.5612.8 (71150)Urea (mg/dl) 71.067.8 (56142) 58.465.8 (2999) 46.065.4 (3163)Creatinine (mg/dl) 1.060.1 (0.71.4) 1.060.1 (0.81.3) 1.160.1 (0.71.4)Total protein (g/dl) 7.860.4 (48.8) 7.160.2 (6.28.6) 7.660.3 (6.99.3)Albumin (g/dl) 2.460.1 (1.53.2) 2.560.1 (23.5) 2.760.1 (2.53.1)Globulin (g/dl) 5.460.3 (2.56.5) 4.660.3 (2.76.2) 4.960.2 (4.36.6)Total bilirubin (mg/dl) 0.360.0 (0.20.5) 0.360.0 (0.10.5) 0.360.0 (0.20.3)Creatine phosphokinase (IU/l) 265.0659.7a (79624) 197.3631.2a (115397) 381.2656.5b (175520)Lactate dehydrogenase (IU/l) 212.6642.8 (55378) 181.1622.7 (79296) 234.2667.9 (78544)Triglycerids (mg/dl) 29.267.7 (475) 36.068.7 (13118) 20.266.0 (6299)


0.2 (2.75.0) 4.26

0.2 (3.25.1) 5.46

0.6 (3.87.8)Uric acid (mmol/l) 0.660.1 (0.10.7) 0.460.1 (0.10.9) 0.360.2 (0.21.2)Sodium (mmol/l) 143.462.3 (140152) 145.461.0 (140152) 144.862.0 (136150)Potassium (mmol/l) 4.460.1 (4.05.4) 4.860.2 (4.05.8) 4.760.2 (4.05.1)Chloride (mmol/l) 111.961.9a (99117) 102.662.0b (96119) 105.562.5ab (99116)Calcium (mmol/l) 2.360.1 (1.82.7) 2.360.1 (1.83.0) 2.160.1 (2.02.5)Phosphorus (mg/dl) 5.560.5 (3.58.2) 5.460.4 (3.97.6) 6.060.7) (3.98.9)Cholinesterase (u/l) 2,320.86203.6a

(8593,385)3,111.96217.4b

(2,1784,294)4,087.06341.2c

(3,2135,109)

a,b Different letters within the same row indicate a difference among groups ( P,0.05).



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differences in hematology and bloodbiochemistry observed among the threelocations are related to the increasedlevels of contact between wolves andhumans/domestic animals in the border

and farmland areas. For example, it ispossible that the lower MCV of wolvesliving around park border compared withthose within the park and farms may bedue to loss of iron associated with bloodloss from external parasites, or, alterna-tively, to a nutritional deficiency (Coles,1984; Failace, 2003; Bain, 2004). Yet, theerythrocyte counts in animals livingaround the park and in farms were higher

than those of individuals living in the parksuggesting that blood loss due to externalparasites may not be the (only) contribut-ing factor to the variations in hematologyamong wolves live in different locations.Thus, further study is required to deter-mine iron status, including serum iron,total iron-binding capacity (i.e., transferrinconcentration), unbound iron-binding ca-pacity, and ferritin concentration (Jain,1993), as well as to identify these specificcauses of the variations in hematology

values among wolves living in differentareas.

It is well known that genetic, physiologic,and pathologic conditions, as well asiatrogenic changes, influence cholinesteraseactivities (Lepage et al., 1985). Exposure toorganophosphate or carbamate insecticidessignificantly decreases cholinesterase activ-ity (Furlanello et al., 2006). Stress also has

been recognized to induce secretion ofacetylcholinesterase, a type of cholinester-ase that modulates hematopoiesis (Grisaruet al., 2001, 2006) and neuronal functions(Small et al., 1996; Nizri et al., 2007), as wellas promoting inflammation associatedthrombopoiesis (Pick et al., 2006). Cholin-esterase has also been suggested to play arole in tumorigenesis, especially in hemo-poietic malignancies (Soreg et al., 1991).

This research is the first study to examineserum total cholinesterase in the manedwolf. We observed that the average serumcholinesterase concentration in maned

wolves (disregarding location) was withinthe range of that reported for the domesticdog (3,405 to 6,561 U/l; Santos et al., 1999;Furlanello et al., 2006). However, when thedata were examined on the basis of location,

we discovered that wolves living on farm-lands consistently had higher cholinesteraseconcentrations than those living near thepark border or within the park. This result issurprising, because we expect that wolvesliving in farmland would have more chanceof exposure to organophosphate/carbonatepesticides commonly used in coffee farmsthan those living inside the park, which inturn would lower cholinesterase activity in

the former population (Furlanello et al.,2006). Therefore, variations in cholinester-ase activity among wolves living in differentlocations may be caused by other factorsassociated with direct or indirect interac-tions with humans and domestic species.Interestingly, the wolves living in farms alsohad higher hematologic values (especiallyerythrocyte count) than those living withinthe protected area. Therefore, our observa-tion of elevated cholinesterase in wolves onfarmlands may have been associated withincreased red blood cell production.

In summary, these data serve as valu-able baseline information on hematologyand serum biochemistry for a largecarnivore living precariously in nature ina protected area as well on adjacent landsunder growing human pressure. Althoughthe maned wolf population associated withthe Serra da Canastra National Park is

generally healthy, there are some clearhematology and serum biochemistry dif-ferences that could be early warning signsthat human development is disturbing (orleading to potential negative influence)animal well health. Thus, the obtaineddata will base the long-term monitoring ofthis population, allowing early detectionfor sanitary problems. Furthermore, anindicator of the real health status of

sampled individuals will be possible dueto an association of the hematologic valuesto the parasitic infections and/or exposureto viral or bacterial agents. This linkage



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finally would allow an effective evaluationof the contribution of geographic locationto the pathogens transmission risks be-tween wild and domestic animals.

ACKNOWLEDGMENTS

This study was funded by Fundo Nacionaldo Meio Ambiente grant 056/03, MorrisAnimal Foundation grant D04ZO-77, theDisney Wildlife Conservation Fund, and theConservation Endowment Fund of the Asso-ciation of Zoo and Aquariums. We are gratefulto Joaquim Maia Neto (Director, ParqueNacional da Serra da Canastra-MG, 20062007) and Vicente de Paula Leite (Director,Parque Nacional da Serra da Canastra-MG,20032006) for support.

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Received for publication 28 January 2008.


may-jr (artigo) hematology and blood chemistry parameters differ in free-ranging maned wolves

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