Hematological and biochemical outcomes of Leishmania infantum infection in the South American coati (Nasua nasua) in an endemic area of visceral leishmaniasis in Central Western Brazil

doi:10.21203/rs.3.rs-8099969/v1

Download PDF

Research Article

Hematological and biochemical outcomes of Leishmania infantum infection in the South American coati (Nasua nasua) in an endemic area of visceral leishmaniasis in Central Western Brazil

https://doi.org/10.21203/rs.3.rs-8099969/v1

This work is licensed under a CC BY 4.0 License

You are reading this latest preprint version

Purpose: Wildlife is a major source of infectious diseases affecting humans and domestic animals; however, the impacts of parasitism on naturally parasitized fauna remain largely unknown. In this study, we evaluated the outcomes of Leishmania infantum infection in blood parameters of South American coatis (Nasua nasua) in an area endemicfor leishmaniasis in the Brazilian Midwest region.

Methods: In total, 128 blood samples were obtained from 77 adult South American coatis. Health status was inferred from hematological and biochemical parameters categorized into the following indicators: red blood cell count, coagulation, immune response, infection response, kidney damage, liver damage (LDI), cardiac damage, skeletal muscle damage (SMDI), nutritional profile, and protein profile (PPI). We compared the hematological and biochemical parameters of infected and uninfected animals using the Student’s t-test and Mann–Whitney U-test, and assessed the direct effects of L. infantum on health indicators and body condition (BC) through path analysis.

Results: Our findings showed that L. infantum infection affected LDI through a positive correlation with alanine and aspartate aminotransferases but had no impact on BC. However, BC was influenced by SMDI and PPI through correlations with creatine kinase and albumin levels, respectively, regardless of parasitism.

Conclusions: Our results indicate that L. infantum may cause long-lasting subclinical infections in coatis living in urban areas of the Brazilian Midwest. We highlight the importance of monitoring the impact of L. infantum infections on wild mammals in leishmaniasis-endemic areas.

Conservation Biology

Health

Procyonidae

Urban Ecology

Anthropogenic environmental changes have increased the emergence of infectious diseases by intensifying the wildlife–human–domestic animal contact, enabling many spillover-spillback events^1–3. This phenomenon is exemplified by the adaptation of Lutzomyia longipalpis, the main vector of Leishmania infantum that historically occurs in rural environments associated with humans and domestic animals (dogs and chickens), to urban environments^4,5. Consequently, L. infantum has become widespread across urbanized areas in Brazil, constituting a public health concern since the early 1980s. Humans and domestic dogs develop severe diseases known as human visceral leishmaniasis (HVL) and canine visceral leishmaniasis (CVL) respectively.

Many wild mammal species have been identified as the reservoir hosts of L. infantum. Indeed, Roque and Jansen and Azami-Conesa et al. listed dozens of wild mammal species parasitized by L. infantum^6,7. Recently de Macedo et al. reported L. infantum infection on the South American coati (Procyonidae, Nasua nasua)⁸ – hereafter referred to as “coati” - in an endemic area for HVL and CVL at Brazilian Midwest.

The effects of parasitism on wild mammals have been reported in previous experimental studies^9–12. Nevertheless, our society's growing awareness of animal experimentation and strict guidelines from ethics committees have limited experimental infections¹³. Although many studies have investigated the consequences of natural infections on free-living animal health have been reported^14–21, only a few reports have evaluated the outcomes of L. infantum on the health of free-living mammals^22–24.

Coatis are medium-sized carnivores widely distributed in South America²⁵. In Brazil, their populations thrive in forested fragments near and within urban areas, possibly due to the absence of natural predators and abundant food supply from anthropogenic sources^26–29, leading them to acquire a synanthropic lifestyle. In Campo Grande (CG), Mato Grosso do Sul (MS), in the Midwest region of Brazil, where HVL and CVL are endemic^30,31, coati populations are present in high densities, are in close proximity to humans²⁹, and may participate in the enzootic cycle of L. infantum⁸.

While most studies on parasites in wild animals are restricted to diagnostic tests aiming to determine the presence or absence of parasitic infections, hematology and serum biochemistry are essential to estimate the impact on their hosts³². Although hematology and serum biochemistry are widely used to monitor the clinical course of HVL and CVL^33–39, only a few studies have explored the consequences of L. infantum infection on wildlife blood parameters^23,40. Therefore, in this study, we aimed to investigate the impact of L. infantum infection on the blood parameters and body condition (BC) of coatis inhabiting forest fragments in CG/MS.

Study areas

This study was performed in two forest fragments in the Brazilian Midwest: (i) a conservation unit called Prosa State Park (20°26’59’’S, 54°33’55’’O), and (ii) a residential area known as Air Force Village (20°28’17’’S, 54°39’14’’O) (Fig. 1).

Capture of coatis and sample collection

Animals were captured and recaptured from March 2018 to May 2019 using box traps (90×45×50; Equipos Fauna®, Brazil), sedated with 7 mg/kg of Telazol anesthetic (Zoetis®, USA), and marked with subcutaneous microchips (Animal Tag®, Brazil). After sedation, animals were clinically inspected, and their dental conditions and body size measurements were recorded to estimate age through model-based analysis⁴¹. Blood was aseptically collected from the femoral vein using 0.25 × 8 mm needles (BD® Vacutainer) and deposited in tubes containing ethylenediamine tetraacetic acid (EDTA), as well as tubes containing clot activator to obtain serum. Bone marrow was aseptically obtained from the manubrium sterni using 0.40 × 1.2 mm hypodermic needles and 10 mL syringes and placed in EDTA tubes. All biological material was kept at 4°C in thermal boxes until laboratory procedures. After complete sedation recovery, animals were released at capture sites. All field procedures were conducted in accordance with the Instituto Chico Mendes de Conservação da Biodiversidade (license number 56912-2), Instituto de Meio Ambiente de Mato Grosso do Sul (IMASUL license number 05/2017, process Nº61/405959/2016), Air Force Cooperation Agreement (Nº01/GAP-CG/2018), and Ethics Committee for Animal Use of Universidade Católica Dom Bosco, CG, MS (license number 001/2017).

Hematological and biochemical analysis

Red blood cell (RBC), white blood cell (WBC), hemoglobin (Hgb), hematocrit (Hct), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) and platelet counts were measured by an automated hematology analyzer (POCH-iV 100, Syxmex^®, Brazil) according to manufacturer’s recommendations. Validation of the Diff pocH-100iV hematological analyzer for use in coatis was performed by a specialized Sysmex technician using blood from five clinically healthy adult coatis together with hematological values for N. nasua reported in the International Species Identification System⁴². Standard quality controls consisted of low, normal, and high levels of animal platelet, RBC, and WBC material (EIGHTCHECK-3WP X-TRA, Sysmex^®), performed daily before each analysis. Different leukocyte counts were performed in blood smears fixed with methanol, stained with Panótico kit (Laborclin^®, Brazil), and visualized using a binocular microscope (Eclipse E200, Nikon^®, Japan). Serum concentrations of creatine kinase (CK), CKMB fraction, lactate dehydrogenase (LDH), alkaline phosphatase (AP), alanine aminotransferase (ALT), aspartate transaminase (AST), urea, creatinine, albumin, total serum protein (TSP), total cholesterol, high-density lipoprotein (HDL) cholesterol, glucose, and triglycerides were determined by spectrophotometer analysis (Brasmed^®, Brazil) using the Biotech^® and Gold^® commercial kits. Globulin levels were determined based on the difference between TSP and albumin levels.

L. infantum detection

Diagnosis of L. infantum infection was based on parasitological, serological, and molecular methods. Bone marrow samples were placed in biphasic media, Novy-MacNeal-Nicolle (NNN), with Schneider’s insect medium overlay supplemented with 10% fetal bovine serum. Tubes were incubated at 26°C–28°C and examined weekly for up to 30 days. Detection of anti-L. infantum IgG and K26/K39 specific antibodies was performed through the enzyme-linked immunosorbent assay (ELISA modified from EIE®, BioManguinhos, Brazil) and the dual path platform chromatographic immunoassay (DPP® CVL Biomanguinhos, Brazil), respectively, as described by de Macedo et al.⁸. Molecular detection of L. infantum was conducted using DNA extracted from bone marrow samples, following the protocol described by de Macedo et al.⁸. DNA samples were first tested by PCR for Leishmania spp. kinetoplastid DNA⁴³. Positive samples were confirmed by PCRs for heat-shock protein 70 (hsp70) and L. infantum kDNA genes^44,45. The hsp70 PCR products were sequenced using the Sanger method on an AB3500 platform (Applied Byosystems®, United States), and the obtained sequences were deposited in GenBank database (accession numbers are described in de Macedo et al.⁸.

Data analysis

To avoid misinterpretation, only adult individuals were included in the analysis, and capture and recapture (at least one month apart) were considered independent. Descriptive statistics (mean ± standard deviation) were used to obtain mean hematological and serum biochemistry values of the uninfected (UN) and infected (LI) animals. A coati was considered infected when its samples were positive in at least one diagnostic method (parasitological, molecular, and serological, if positive in both ELISA and DPP®CVL), as described in de Macedo et al.⁸. The Shapiro–Wilk test was used to check the normality of the hematological and biochemical parameters, and the Student’s t-test and Mann–Whitney U-test were performed according to data distribution to observe differences between the UN and LI groups.

To determine the indirect impact of L. infantum infection on BC through health indicators, we performed path analysis according to Santos et al.¹⁸. We estimated BC based on residuals from an ordinary linear regression between body weight and head-to-body length of the coatis. Health indicators were categorized into: (i) red blood cell (RBCI) (RBC, Hgb, Hct, MCV, and MCHC); (ii) coagulation (COI) (platelets); (iii) immune response (IMRI) (globulin and lymphocytes); (iv) infection response (IRI) (monocytes, neutrophils and eosinophils); (v) kidney damage (KDI) (urea and creatinine); (vi) liver damage (LDI) (AST, ALT, and AP); (vii) cardiac damage (CDI) (CKMB); (viii) skeletal muscle damage (SMDI) (CK and LDH); (ix) nutritional profile (NPI) (triglycerides, total cholesterol, HDL cholesterol, and glucose); and (x) protein profile (PPI) (albumin and TSP). Variables were considered statistically significant for p values < 0.05. All data were analyzed using R software⁴⁶.

A total of 128 samples were obtained from 77 adult coatis. Forty-nine samples (38.3%) were positive: 33 (67.3%) positive by serology, 9 (18.4%) by molecular methods, and 7 (14.3%) by both methods. No samples were positive in the bone marrow aspirate, and 79 samples (61.7%) were negative using all diagnostic methods. When evaluating recaptures (n = 51), we observed three individuals that were positive at first capture and negative at the subsequent recapture.

In the clinical evaluation, the most frequent findings were wounds (32.8%), paw pad hyperkeratosis (31.2%), and scars (27.3%), and 34 (26.6%) animals did not present any clinical findings (Online Resource 1). The clinical signs suggestive of CVL as lymphadenopathy (7.8%; UN = 8.9%, LI = 6.1%), alopecia (3.1%; UN = 3.8%, LI = 2%), and desquamation (2.3%; UN = 2.5%, LI = 2%) were less frequent and mostly observed in uninfected animals, including paw pad hyperkeratosis (UN = 39.2%, LI = 18.4%).

Despite high standard deviations in some hematological and serum biochemical parameters, we present the mean comparison test results in Table 1; however, biological interpretation of these results must be carried out with caution. Mean comparison tests showed no significant differences in hematological parameters between the UN and LI groups. In contrast, regarding serum biochemistry, we observed that the LI group presented higher ALT (p = 0.01), AST (p = 0.04), and HDL cholesterol (p = 0.02) levels than the UN group (Table 1).

Table 1
Hematological and serum biochemistry parameters of South American coatis (*Nasua nasua*) uninfected and infected by *Leishmania infantum* at Campo Grande, Mato Grosso do Sul, Brazil. Values are expressed by mean ± standard deviation.
Parameters	Uninfected (n = 79)	L. infantum infected (n = 49)	p value	Statistical test
Hematology
RBC (10x⁶/mm³)	5.39 ± 0.50	5.39 ± 0.56	0.99	Student’s T
Hgb (g/dL)	10.50 ± 0.98	10.50 ± 1.07	0.98	Student’s T
Hct (%)	33.78 ± 3.19	34.08 ± 3.28	0.62	Student’s T
MCV (fL)	59.68 ± 2.58	59.86 ± 3.43	0.76	Student’s T
MCHC (g/dL)	32.66 ± 0.99	32.62 ± 1.07	0.80	Mann-Whitney U
Platelet (10x³/mm³)	550582.30 ± 152968.90	582857.10 ± 168919.90	0.42	Mann-Whitney U
WBC (10x³/mm³)	16003.80 ± 4889.27	16995.92 ± 5442.58	0.25	Mann-Whitney U
Neutrophil (10x³/mm³)	10856.77 ± 5365.28	12159.90 ± 5922.79	0.26	Mann-Whitney U
Lymphocyte (10x³/mm³)	3131.89 ± 2163.59	2480.14 ± 1556.47	0.10	Mann-Whitney U
Monocyte (10x³/mm³)	454.99 ± 386.44	654.88 ± 864.12	0.32	Mann-Whitney U
Eosinophil (10x³/mm³)	1538.19 ± 1068.90	1292.76 ± 1054.79	0.10	Mann-Whitney U
Basophil (10x³/mm³)	21.96 ± 57.02	12.22 ± 53.27	0.18	Mann-Whitney U
Serum biochemistry
CK (U/L)	1792.96 ± 1705.03	1921.80 ± 1555.73	0.29	Mann-Whitney U
CKMB (IU/L)	2092.03 ± 957.51	2127.84 ± 894.82	0.84	Mann-Whitney U
LDH (IU/L)	1528.68 ± 786.09	1641.84 ± 754.99	0.26	Mann-Whitney U
AST (IU/L)	194.97 ± 123.95	231.97 ± 130.43	0.04	Mann-Whitney U
ALT (IU/L)	96.21 ± 51.70	116.18 ± 52.46	0.01	Mann-Whitney U
AP (IU/L)	26.28 ± 16.78	21.10 ± 8.21	0.11	Mann-Whitney U
Urea (mmol/L)	26.49 ± 15.43	32.28 ± 53.44	1.00	Mann-Whitney U
Creatinine (µmol/L)	1.23 ± 0.54	1.12 ± 0.33	0.44	Mann-Whitney U
Albumin (g/dL)	1.97 ± 1.11	2.28 ± 1.33	0.27	Mann-Whitney U
Globulin (g/dL)	5.48 ± 1.49	5.62 ± 1.43	0.60	Student’s T
TSP (g/dL)	7.45 ± 1.58	7.90 ± 1.62	0.05	Mann-Whitney U
Total cholesterol (mmol/L)	153.61 ± 118.50	160.73 ± 123.67	0.85	Mann-Whitney U
HDL cholesterol (mmol/L)	57.25 ± 39.94	68.49 ± 38.80	0.02	Mann-Whitney U
Glucose (mmol/L)	85.27 ± 31.08	89.37 ± 33.60	0.60	Mann-Whitney U
Triglycerides (mmol/L)	48.29 ± 53.59	59.54 ± 77.19	0.20	Mann-Whitney U

ALT: Alanine aminotransferase; AP: Alkaline phosphatase; AST: Aspartate aminotransferase; CK: Creatine kinase; CKMB: Creatine kinase (MB fraction); Hct: Hematocrit; Hgb: Hemoglobin; LDH: Lactate dehydrogenase; MCHC: Mean corpuscular hemoglobin concentration; MCV: Mean corpuscular volume; RBC: Red blood cells; TSP: Total serum protein; WBC; White blood cells.

Path analysis showed that L. infantum directly affected LDI (path coefficient = 0.23, p < 0.05) through positive correlation with ALT (r = 0.92) and AST (r = 0.67) but did not impact BC (Fig. 2). Additionally, we observed positive correlations between BC and SMDI through CK (path coefficient = -0.18, p < 0.05, r = -0.81) and PPI through albumin (path coefficient = -0.35, p < 0.05, r = -0.98), irrespective of L. infantum infection (Fig. 2).

Our data analysis indicated that infection by L. infantum in coatis inhabiting urban forested areas in CG may result in hepatic damage, as demonstrated by the observed positive correlation between infection and ALT and AST levels. This was reinforced by higher mean ALT and AST values in LI individuals than in UN individuals. These aminotransferases are predominantly hepatocellular, indicating both acute and chronic liver injuries when detected at high serum concentrations^47,48. Indeed, elevated levels of ALT and AST have been correlated with hepatic inflammatory processes during visceral leishmaniasis (VL)^49,50 and with clinical signs and progression of both CVL and HVL^39,51–58. Nevertheless, these findings may also relate to many other factors, since the sampled coatis were free-living animals exposed to several conditions in the urban environment, including consumption of discarded food in trash cans^29,59, and other infectious agents^60,61.

Although L. infantum infection was positively correlated with LDI, it did not appear to affect the health status of the infected coatis because we found no influence on BC, and no significant difference in mean comparison tests of biochemical parameters related to the nutritional profile, such as triglycerides, total cholesterol, and glucose, between the UN and LI groups. Since young HVL patients present changes in weight and body mass^62–64, we may not have observed the impact of L. infantum infection on BC because only adult individuals were included in the analysis.

The higher HDL cholesterol level observed in the LI group suggests a chronic infection phase at the time of capture, because serum HDL cholesterol concentrations are reported to be low in acute symptomatic infections in both CVL and HVL^{34,35,57,65,66}. In Leishmania spp. infections, HDL cholesterol has been positively correlated with gamma-interferon (IFN-γ) and negatively with interleukin 10 (IL-10), cytokines related to protection and progression of VL respectively^67,68. In fact, while the Th1 response, mediated by IFN-γ, prevails in asymptomatic infections, the Th2 response, mediated by some cytokines including IL-10, is more prevalent in acute symptomatic infections by Leishmania spp.^67,69,70. Additionally, since serum concentration of HDL cholesterol is reported to be inversely correlated with amastigote load in lymphoid tissues in HVL^71,72, our results may indicate that the sampled coati populations possess physiological protective mechanisms against L. infantum infection. High serum HDL cholesterol levels impair Leishmania spp. internalization across the macrophage plasma membrane^73–75. Moreover, we observed fewer infected coatis showing clinical signs compatible with CVL than the coatis in the UN group (Online Resource 1).

Path analysis conducted in this study indicated that other factors, irrespective of L. infantum infection, might influence the BC of coatis. We observed that PPI influenced BC through positive correlation with albumin levels (Fig. 2), which may be explained by the positive association between serum albumin and muscle mass^76–78. Although increased serum protein levels may relate to dehydration, this is usually accompanied by an increase in Hct⁷⁹, which was not observed in our study, and dehydration was not observed in the sampled coatis during the capture events.

SMDI influenced BC through positive correlation with CK, an indirect marker related to muscle membrane disruption, and inflammatory response as a consequence of metabolic demands during physical effort⁸⁰. As skeletal muscle is the most important source of CK, a higher body mass might increase the susceptibility to mechanical load-induced muscle membrane damage^80,81, leading to greater CK release into the bloodstream. Therefore, we predict that the capture method used in the present study may be more harmful to coatis with higher BC.

Since host-parasite coevolution has drastically changed due to human encroachment of natural environments, continuous monitoring of wildlife health in urbanized areas is extremely important for the conservation of biodiversity and better understanding of the parasite ecology. As the host’s physiological state influences parasite transmission, health status assessment provides important data for monitoring emerging and reemerging diseases in urban areas.

This study presented the effects of L. infantum infection on the health of free-living coati in an urban environment, as expressed by hematological, biochemical, and BC parameters. Our results demonstrate that L. infantum infection may lead to alterations in liver function and lipid profile of infected coatis but may not result in clinical disease presentation. This study highlights the importance of conservation of one of the most common wild carnivorous species inhabiting urban forest fragments in Brazil, from a One Health perspective, given the zoonotic and endemic characteristics of L. infantum in the Brazilian Midwest.

Author contributions

GCM, ALRR, CEO, WTGB and HMH conceived and designed the study. GCM, WTGB, WOA, ACR and JGBP performed the research. GCM, FMS, NYS and SCCX analyzed the data. GCM, WTGB, and ARS, drafted the initial manuscript. ALRR, GEOP, GBA, AMJ and HMH reviewed and edited the original draft. All the authors approved the final version of the manuscript.

Funding

GCM was supported by the Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (project number SEI E-26/200.295/2025), and ALRR and HMH were supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (project numbers 309507/2023-5 and 311769/2023-3 respectively).

Acknowledgements

The authors are grateful to the Air Force Village and the Prosa State Park staffs for the cooperation with the field works, and to the laboratory LABDOC for the support in the hematological and biochemical analyses.

Declaration of conflict of interest

The authors declare no conflict of interests.

Bradley CA, Altizer S (2007) Urbanization and the ecology of wildlife diseases. Trends Ecol Evol 22(2):95-102. https://doi.org/10.1016/j.tree.2006.11.001
Acevedo-Whitehouse K, Duffus AL (2009) Effects of environmental change on wildlife health. Philos Trans R Soc Lond B Bio Sci 364(1534):3429-38. https://doi.org/10.1098/rstb.2009.0128
Hassell JM, Begon M, Ward MJ, Fèvre EM (2017) Urbanization and Disease Emergence: Dynamics at the Wildlife-Livestock-Human Interface. Trends Ecol Evol 32(1):55-67. https://doi.org/10.1016/j.tree.2016.09.012
Werneck GL (2008) Forum: geographic spread and urbanization of visceral leishmaniasis in Brazil. Introduction. Cadernos De Saúde Pública 24(12):2937–40. https://doi.org/10.1590/S0102-311X2008001200023
de Souza Fernandes W, Infran JOM, de Oliveira EF, Casaril AE, Barrios SPG, de Oliveira SLL, et al (2022) Phlebotomine sandfly (Diptera: Psychodidae) fauna and the association between climatic variables and the abundance of Lutzomyia longipalpis sensu lato in an intense transmission area for visceral leishmaniasis in Central Western Brazil. J Med Entomol 59(3):997-1007. https://doi.org/10.1093/jme/tjac006
Roque AL, Jansen AM (2014) Wild and synanthropic reservoirs of Leishmania species in the Americas. Int J Parasitol Parasites Wildl 3(3):251-62. https://doi.org/10.1016/j.ijppaw.2014.08.004
Azami-Conesa I, Gómez-Muñoz MT, Martínez-Díaz RA (2021) A Systematic Review (1990-2021) of Wild Animals Infected with Zoonotic Leishmania. Microorganisms9(5):1101. https://doi.org/10.3390/microorganisms9051101
de Macedo GC, Barreto WTG, de Oliveira CE, Santos FM, Porfírio GEO, Xavier SCDC, et al (2023a) Leishmania infantum infecting the carnivore Nasua nasua from urban forest fragments in an endemic area of visceral leishmaniasis in Brazilian Midwest. Front Cell InfectMicrobiol 12:1050339. https://doi.org/10.3389/fcimb.2022.1050339
Lindsay DS, Hendrix CM, Blagburn BL (1988) Experimental Cryptosporidium parvum infections in opossums (Didelphis virginiana). J Wildl Dis 24(1):157-9. https://doi.org/10.7589/0090-3558-24.1.157
Franke CR, Greiner M, Mehlitz D (1994) Monitoring of clinical, parasitological and serological parameters during an experimental infection of capybaras (Hydrochaeris hydrochaeris) with Trypanosoma evansi. Acta Trop 58(2):171-4. https://doi.org/10.1016/0001-706x(94)90056-6
Araujo Carreira JC, Jansen AM, Deane MP, Lenzi HL (1996) Histopathological study of experimental and natural infections by Trypanosoma cruzi in Didelphis marsupialis. Mem Inst Oswaldo Cruz 91(5):609-18. https://doi.org/10.1590/s0074-02761996000500012
Herrera HM, Aquino LP, Menezes RF, Marques LC, Moraes MA, Werther K, et al (2001) Trypanosoma evansi experimental infection in the South American coati (Nasua nasua): clinical, parasitological and humoral immune response. Vet Parasitol 102(3):209-16. https://doi.org/10.1016/s0304-4017(01)00532-5
Singh VP, Pratap K, Sinha J, Desiraju K, Bahal D, Kukreti R (2016) Critical evaluation of challenges and future use of animals in experimentation for biomedical research. Int J Immunopathol Pharmacol 29(4):551-61. https://doi.org/10.1177/0394632016671728
Ruykys L, Rich B, McCarthy P (2012) Haematology and biochemistry of warru (Petrogale lateralis Mac-Donnell Ranges race) in captivity and the wild. Aust Vet J 90(9):331–40. https://doi.org/10.1111/j.1751-0813.2012.00956.x
Clarke J, Warren K, Calver M, de Tores P, Mills J, Robertson, I (2013) Hematologic and serum biochemical reference ranges and assessment of exposure to infectious diseases prior to translocation of the threatened Western ringtail opossum (Pseudocheirus occidentalis). J Wildl Dis 49(4):831–40. https://doi.org/10.7589/2011-12-345
Pacioni C, Robertson ID, Maxwell M, van Weenen J, Wayne AF (2013) Hematologic characteristics of the woylie (Bettongia penicillata ogilbyi). J Wildl Dis 49(4):816–30. https://doi.org/10.7589/2011-09-275
Olifiers N, Jansen AM, Herrera HM, Bianchi R de C, D'Andrea PS, Mourão G de M, et al (2015) Coinfection and Wild Animal Health: Effects of Trypanosomatids and Gastrointestinal Parasites on Coatis of the Brazilian Pantanal. PLoS One 10(12):e0143997. https://doi.org/10.1371/journal.pone.0143997
Santos FM, de Macedo GC, Barreto WTG, Oliveira-Santos LGR, Garcia CM, Mourão GM, et al (2018) Outcomes of Trypanosoma cruzi and Trypanosoma evansi infections on health of South American coati (Nasua nasua), crab-eating fox (Cerdocyon thous), and ocelot (Leopardus pardalis) in the Brazilian Pantanal. PLoS One 13(8):e0201357. https://doi.org/10.1371/journal.pone.0201357
Nantes WAG, Barreto WTG., Santos FM., de Macedo GC, Rucco, AC, Assis WO, et al (2019) The influence of parasitism by Trypanosoma cruzi in the hematological parameters of the white ear opossum (Didelphis albiventris) from Campo Grande, Mato Grosso do Sul, Brazil. Int J Parasitol Parasites Wildl 9:16-20. https://doi.org/10.1016/j.ijppaw.2019.03.015
Santos FM, de Macedo GC, Barreto WTG, Nantes WAG, de Assis WO, Herrera HM (2019) Hematological values of crab-eating foxes (Cerdocyon thous) from Pantanal, Mato Grosso do Sul, Brazil, naturally infected and non-infected by Trypanosoma cruzi e T. evansi. Ciênc Anim Bras 20:e-50604. https://doi.org/10.1590/1089-6891v20e-50604
da Silva AR, Herrera HM, de Oliveira CE, Torres JM, Ferreira AMR, Leite JDS, et al (2023) The relationships among Leishmania infantum and phyllostomid bats assessed by histopathological and molecular assays. Int J Parasitol Parasites Wildl 23:100904. https://doi.org/10.1016/j.ijppaw.2023.100904
Courtenay O, Quinnell RJ, Garcez LM, Dye C (2002) Low infectiousness of a wildlife host of Leishmania infantum: the crab-eating fox is not important for transmission. Parasitol 125(Pt 5):407-14. https://doi.org/10.1017/s0031182002002238
Millán J, Chirife AD, Altet L (2015) Serum chemistry reference values for the common genet (Genetta genetta): variations associated with Leishmania infantum infection. Vet Q 35(1):43-7. https://doi.org/10.1080/01652176.2014.987883
da Silva MAD, Oliveira MR, Schettino SC, dos Santos IG, Neto MBO, da Silva WSI, et al (2021) New insights on severe clinical manifestations and deaths from visceral leishmaniasis in free-living crab-eating foxes (Cerdocyon thous) in Brazil. Res Soc Devel 10(16):e108101622869. https://doi.org/10.33448/rsd-v10i16.22869
Gompper ME, Decker DM (1998) Nasua nasua. Mamm Species 580: 1–9
Bovendorp R, Galetti M (2007) Density and population size of mammals introduced on a land-bridge island in southeastern Brazil. Biol Invasions 9:353–7. https://doi.org/10.1007/s10530-006-9031-7
Costa EMJ, Mauro RA, Silva JSV (2009) Group composition and activity patterns of brown-nosed coatis in savanna fragments, Mato Grosso do Sul, Brazil. Braz J Biol 69(4):985-91. https://doi.org/10.1590/S1519-69842009000500002
Estevam LGTM, Fonseca Junior AA, Silvestre BT, Hemetrio NS, Almeida LR, Oliveira MM, et al (2020) Seven years of evaluation of ectoparasites and vector-borne pathogens among ring-tailed coatis in an urban park in southeastern Brazil. Vet Parasitol Reg Stud Reports 21:100442. https://doi.org/10.1016/j.vprsr.2020.100442
Barreto WTG, Herrera HM., de Macedo, GC, Rucco AC, Santos LGO, de Assis WO, et al (2021) Density and survivorship of the South American coati (Procyonidae: Nasua nasua) in the urban area of Campo Grande, Centra-Western Brazil. Hystrix It J Mamm 32(1):82-8. https://doi.org/10.4404/hystrix-00386-2020
Brazuna JCM, Araujo e Silva E, Brazuna JM, Domingos IH, Chaves N, Honer MR, et al (2012) Profile and geographic distribution of reported cases of visceral leishmaniasis in Campo Grande, State of Mato Grosso do Sul, Brazil, from 2002 to 2009. Rev Soc Bras Med Trop 45(5):601-6. https://doi.org/10.1590/S0037-86822012000500012
de Sousa KC, André MR, Herrera HM, de Andrade GB, Jusi MM, dos Santos LL, et al (2013) Molecular and serological detection of tick-borne pathogens in dogs from an area endemic for Leishmania infantum in Mato Grosso do Sul, Brazil. Rev Bras Parasitol Vet 22(4):525-31. https://doi.org/10.1590/S1984-29612013000400012
Bernal-Valle S, Teixeira MN, de Araújo Neto AR, Gonçalves-Souza T, Feitoza BF, dos Santos SM, et al (2022) Parasitic infections, hematological and biochemical parameters suggest appropriate health status of wild coati populations in anthropic Atlantic Forest remnants. Vet Parasitol Reg Stud Reports 30:100693. https://doi.org/10.1016/j.vprsr.2022.100693
Reis AB, Martins-Filho OA, Teixeira-Carvalho A, Carvalho MG, Mayrink W, França-Silva JC, et al (2006) Parasite density and impaired biochemical/hematological status are associated with severe clinical aspects of canine visceral leishmaniasis. Res Vet Sci 81(1):68-75. https://doi.org/10.1016/j.rvsc.2005.09.011
de Freitas JC, Nunes-Pinheiro DC, Lopes Neto BE, Santos GJ, Abreu CR, Braga RR, et al (2012) Clinical and laboratory alterations in dogs naturally infected by Leishmania chagasi. Rev Soc Bras Med Trop 45(1):24-9. https://doi.org/10.1590/s0037-86822012000100006
Gatto M, de Abreu MM, Tasca KI, Simão JC, Fortaleza CM, Pereira PC, et al (2013) Biochemical and nutritional evaluation of patients with visceral leishmaniasis before and after treatment with leishmanicidal drugs. Rev Soc Bras Med Trop 46(6):735-40. https://doi.org/10.1590/0037-8682-0198-201
Ribeiro RR, da Silva SM, Fulgêncio Gde O, Michalick MS, Frézard FJ (2013) Relationship between clinical and pathological signs and severity of canine leishmaniasis. Rev Bras Parasitol Vet 22(3):373-8. https://doi.org/10.1590/S1984-29612013000300009
Montargil SMA, Carvalho FS, de Oliveira GMS, Munhoz AD, Alberto Carlos RS, Wenceslau AA (2018) Clinical, Hematological and Biochemical Profiles of Dogs with Leishmania infantum. Acta Scientiae Veterinariae 46(1):7. https://doi.org/10.22456/1679-9216.82065
Silva JL, Oliveira VVG, Silva LAMT, Silva RPE, Alves LC, Cavalcanti MP, et al (2019) Evaluation of serum biochemical parameters, structural changes, immunohistochemistry and parasite load in the urinary system of dogs infected naturally by Leishmania infantum. J Comp Pathol 167:26-31. https://doi.org/10.1016/j.jcpa.2018.11.007
Tesfanchal B, Gebremichail G, Belay G, Gebremariam G, Teklehaimanot G, Haileslasie H, et al (2020) Alteration of Clinical Chemistry Parameters Among Visceral Leishmaniasis Patients in Western Tigrai, Ethiopia, 2018/2019: A Comparative Cross-Sectional Study. Infect Drug Resist 13:3055-62. https://doi.org/10.2147/IDR.S261698
Cavalera MA, Iatta R, Laricchiuta P, Passantino G, Abramo F, Mendoza-Roldan JA, et al (2020) Clinical, haematological and biochemical findings in tigers infected by Leishmania infantum. BMC Vet Res 16(1):214. https://doi.org/10.1186/s12917-020-02419-y
Olifiers N, Bianchi RC, D’Andrea PS, Mourão G, Gomppers ME (2010) Estimating age of carnivores from the Pantanal region of Brazil. Wildl Biol 16:389–9. https://doi.org/10.2981/09-104
International Species Identification System - ISIS (2002). Reference ranges for physiological values in captive wildlife. Apple Valley
Schubach A, Haddad F, Oliveira-Neto MP, Degrave W, Pirmez C, Grimaldi G Jr, et al (1998) Detection of Leishmania DNA by polymerase chain reaction in scars of treated human patients. J Infect Dis 178(3):911–4. https://doi.org/10.1086/515355
Cortes S, Rolão N, Ramada J, Campino L (2004) PCR as a rapid and sensitive tool in the diagnosis of human and canine leishmaniasis using Leishmania donovani s.l.-specific kinetoplastid primers. Trans R Soc Trop Med Hyg 98(1):12-7. https://doi.org/10.1016/s0035-9203(03)00002-6
Graça GC, Volpini AC, Romero GA, Oliveira Neto MP, Hueb M, Porrozzi R, et al (2012) Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species. Mem Inst Oswaldo Cruz 107(5):664–74. https://doi.org/10.1590/s0074-02762012000500014
R Core Team. R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria; 2023 [updated 2025 June 13; cited 2025 June 18]. Available from: http://www.R-project.org/
Giannini EG, Testa R, Savarino V (2005) Liver enzyme alteration: a guide for clinicians. CMAJ 172(3):367-79. https://doi.org/10.1503/cmaj.1040752
Lidbury JA, Steiner JM (2013) Liver-diagnostic evaluation. In: Washabau RJ, Day J (eds)Canine & feline gastroenterology. Elsevier, St. Louis, pp 863-75
Duarte MI, Corbett CE (1987) Histopathological patterns of the liver involvement in visceral leishmaniasis. Rev Inst Med Trop Sao Paulo 29:131–6. https://doi.org/10.1590/s0036-46651987000300003
Kausalya S, Malla N, Ganguly NK, Mahajan RC (1993) Leishmania donovani: in vitro evidence of hepatocyte damage by Kupffer cells and immigrant macrophages in a murine model. Exp Parasitol 77(3):326-33. https://doi.org/10.1006/expr.1993.1090
el Hag IA, Hashim FA, el Toum IA, Homeida M, el Kalifa M, el Hassan AM (1994) Liver morphology and function in visceral leishmaniasis (Kala-azar). J Clin Pathol 47(6):547-51. https://doi.org/10.1136/jcp.47.6.547
Ciaramella P, Oliva G, Luna RD, Gradoni L, Ambrosio R, Cortese L, et al (1997) A retrospective clinical study of canine leishmaniasis in 150 dogs naturally infected by Leishmania infantum. Vet Rec 141(21):539‐43. https://doi.org/10.1136/vr.141.21.539
Amusategui I, Sainz A, Rodríguez F, Tesouro MA (2003) Distribution and relationships between clinical and biopathological parameters in canine leishmaniasis. Eur J Epidemiol 18(2):147-56. https://doi.org/10.1023/a:1023090929302
Giunchetti RC, Mayrink W, Carneiro CM, Corrêa-Oliveira R, Martins-Filho OA, Marques MJ, et al (2008) Histopathological and immunohistochemical investigations of the hepatic compartment associated with parasitism and serum biochemical changes in canine visceral leishmaniasis. Res Vet Sci 84(2):269‐77. https://doi.org/10.1016/j.rvsc.2007.04.020
Mathur P, Samantaray JC, Samanta P (2008) High prevalence of functional liver derangement in visceral leishmaniasis at an Indian tertiary care center. Clin Gastroenterol Hepatol 6(10): 1170‐2. https://doi.org/10.1016/j.cgh.2008.04.033
Oliveira JM, Fernandes AC, Dorval MEC, Alves TP, Fernandes TD, Oshiro ET (2010) Mortality due to visceral leishmanisasis: clinical and laboratory characteristics. Rev Soc Bras Med Trop 43(2): 188-93. https://doi.org/10.1590/s0037-86822010000200016
Heidarpour M, Soltani S, Mohri M, Khoshnegah J (2012) Canine visceral leishmaniasis: relationships between oxidative stress, liver and kidney variables, trace elements, and clinical status. Parasitol Res 111(4): 1491-6. https://doi.org/10.1007/s00436-012-2985-8
Lima IS, Silva JS, Almeida VA, Junior FG, Souza PA, Larangeira DF, et al (2014) Severe clinical presentation of visceral leishmaniasis in naturally infected dogs with disruption of the splenic white pulp. PLoS One 9(2): e87742. https://doi.org/10.1371/journal.pone.0087742
de Macedo GC, Barreto WTG, de Assis WO, Rucco AC, Santos FM, Porfírio GEO, et al (2023b) Hematology and biochemistry of South American coatis Nasua nasua (Carnivora: Procyonidae) inhabiting urban fragments in Midwest Brazil: differences according to intrinsic features and sampling site. Eur J Wildl Res 69:119. https://doi.org/10.1007/s10344-023-01753-4
Perles L, Barreto WTG, Santos FM, Duarte LL, de Macedo GC, Barros-Battesti DM, et al (2023a). Molecular Survey of Hemotropic Mycoplasma spp. and Bartonella spp. in Coatis (Nasua nasua) from Central-Western Brazil. Pathogens 12(4):538. https://doi.org/10.3390/pathogens12040538
Perles L, Barreto WTG, de Macedo GC, Calchi AC, Bezerra-Santos M, Mendoza-Roldan JA, et al (2023b) Molecular detection of Babesia spp. and Rickettsia spp. in coatis (Nasua nasua) and associated ticks from midwestern Brazil. Parasitol Res 122(5):1151-8. https://doi.org/10.1007/s00436-023-07815-5
Zijlstra EE, Ali MS, el-Hassan AM, el-Toum IA, Satti M, Ghalib HW (1992) Clinical aspects of kala-azar in children from the Sudan: a comparison with the disease in adults. J Trop Pediatr 38(1):17-21. https://doi.org/10.1093/tropej/38.1.17
Caldas AJ, Costa J, Aquino D, Silva AA, Barral-Netto M, Barral A (2006) Are there differences in clinical and laboratory parameters between children and adults with American visceral leishmaniasis? Acta Trop 97(3):252-8. https://doi.org/10.1016/j.actatropica.2005.09.010
Maciel BL, Lacerda HG, Queiroz JW., Galvão J, Pontes NN, Dimenstein R, et al (2008) Association of nutritional status with the response to infection with Leishmania chagasi. Am J Trop Med Hyg 79(4):591-8. https://doi.org/10.4269/ajtmh.2008.79.591
Nieto CG, Barrera R, Habela MA, Navarrete I, Molina C, Jiménez A, et al (1992) Changes in the plasma concentrations of lipids and lipoprotein fractions in dogs infected with Leishmania infantum. Vet Parasitol 44(3-4):175-82. https://doi.org/10.1016/0304-4017(92)90115-p
Soares NM, Leal TF, Fiúza MC, Reis EA, Souza MA, Dos-Santos WL, et al (2010) Plasma lipoproteins in visceral leishmaniasis and their effect on Leishmania-infected macrophages. Parasite Immunol 32(4):259-66. https://doi.org/10.1111/j.1365-3024.2009.01187.x
Caldas A, Favali C, Aquino D, Vinhas V, van Weyenbergh J, Brodskyn C, et al (2005) Balance of IL-10 and interferon-gamma plasma levels in human visceral leishmaniasis: implications in the pathogenesis. BMC Infect Dis 5:113. https://doi.org/10.1186/1471-2334-5-113
Paswan RK, Bimal S, Kumari A, Sinha P, Rabidas VN, Pandey K, et al (2016) Reduced high density lipoprotein concentration modulates increased interleukin-10 and decreased interferon-gamma in Visceral Leishmaniasis patients. Gen Med 4(2):1000233. https://doi.org/10.4172/2327-5146.1000233
Carvalho EM, Badaró R, Reed SG, Jones TC, Johnson WD Jr (1985) Absence of gamma interferon and interleukin 2 production during active visceral leishmaniasis. J Clin Invest 76(6):2066-9. https://doi.org/10.1172/JCI112209
Ghalib HW, Piuvezam MR, Skeiky YA, Siddig M, Hashim FA, el-Hassan AM (1993) Interleukin 10 production correlates with pathology in human Leishmania donovani infections. J Clin Invest 92(1):324-9. https://doi.org/10.1172/JCI116570
Lal CS, Verma N, Rabidas VN, Ranjan A, Pandey K, Verma RB, et al (2010) Total serum cholesterol determination can provide understanding of parasite burden in patients with visceral leishmaniasis infection. Clin Chim Acta 411:2112-3. https://doi.org/10.1016/j.cca.2010.08.041
Ghosh J, Lal CS, Pandey K, Das VN, Das P, Roychoudhury K, et al (2011) Human visceral leishmaniasis: decrease in serum cholesterol as a function of splenic parasite load. Ann Trop Med Parasitol 105(3):267-71. https://doi.org/10.1179/136485911X12899838683566
Pucadyil TJ, Tewary P, Madhubala R, Chattopadhyay A (2004) Cholesterol is required for Leishmania donovani infection: implications in leishmaniasis. Mol Biochem Parasitol 133(2):145-52. https://doi.org/10.1016/j.molbiopara.2003.10.002
Norata GD, Pirillo A, Ammirati E, Catapano AL (2012) Emerging role of high-density lipoproteins as a player in the immune system. Atherosclerosis 220(1):11-21. https://doi.org/10.1016/j.atherosclerosis.2011.06.045
Kumar GA, Roy S, Jafurulla M, Mandal C, Chattopadhyay A (2016) Statin-induced chronic cholesterol depletion inhibits Leishmania donovani infection: Relevance of optimum host membrane cholesterol. Biochim Biophys Acta 1858(9):2088-96. https://doi.org/10.1016/j.bbamem.2016.06.010
Baumgartner RN, Koehler KM, Romero L, Garry PJ (1996) Serum albumin is associated with skeletal muscle in elderly men and women. Am J Clin Nutr 64(4):552-8. https://doi.org/10.1093/ajcn/64.4.552
Snyder CK, Lapidus JA, Cawthon PM, Dam TT, Sakai LY, Marshall LM, et al (2012) Serum albumin in relation to change in muscle mass, muscle strength, and muscle power in older men. J Am Geriatr Soc 60(9):1663-72. https://doi.org/10.1111/j.1532-5415.2012.04115.x
Kim H, Suzuki T, Kim M, Kojima N, Yoshida Y, Hirano H, et al (2015) Incidence and predictors of sarcopenia onset in community-dwelling elderly Japanese women: 4-year follow-up study. J Am Med Dir Assoc 16(1):85.e1-8. https://doi.org/10.1016/j.jamda.2014.10.006
Abutarbush S (2015) Hematological and serum biochemical findings in clinical cases of cattle naturally infected with lumpy skin disease. J Infect Dev Ctries 9(3):283-8. https://doi.org/10.3855/jidc.5038
Brancaccio P, Lippi G, Maffulli N. Biochemical markers of muscular damage (2010) Clin Chem Lab Med 48(6):757-67. https://doi.org/10.1515/CCLM.2010.179
Knoblauch MA, O'Connor DP, Clarke MS (2013) Obese mice incur greater myofiber membrane disruption in response to mechanical load compared with lean mice. Obesity (Silver Spring) 21(1):135-43. https://doi.org/10.1002/oby.20253

No competing interests reported.

ESM1.xlsx.xlsx

Download PDF

Editorial decision: Revision requested
01 Dec, 2025
Reviews received at journal
30 Nov, 2025
Reviews received at journal
21 Nov, 2025
Reviewers agreed at journal
18 Nov, 2025
Reviewers agreed at journal
17 Nov, 2025
Reviewers invited by journal
13 Nov, 2025
Editor assigned by journal
13 Nov, 2025
Submission checks completed at journal
13 Nov, 2025
First submitted to journal
12 Nov, 2025

You are reading this latest preprint version

Hematological and biochemical outcomes of Leishmania infantum infection in the South American coati (Nasua nasua) in an endemic area of visceral leishmaniasis in Central Western Brazil

Status:

Version 1

Abstract

Figures

Introduction

Materials and Methods

Study areas

Capture of coatis and sample collection

Hematological and biochemical analysis

L. infantum detection

Data analysis

Results

Discussion

Conclusions

Declarations

References

Additional Declarations

Supplementary Files

Status:

Version 1